Salts of a pim kinase inhibitor

ABSTRACT

The present invention relates to salt forms of the Pim kinase inhibitor N-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide, including methods of preparation thereof, and intermediates in the preparation thereof, where the compound is useful in the treatment of Pim kinase-related diseases such as cancer.

FIELD OF THE INVENTION

The present invention relates to salt forms of the Pim kinase inhibitorN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide,including methods of preparation thereof, and intermediates in thepreparation thereof, where the compound is useful in the treatment ofPim kinase-related diseases such as cancer.

BACKGROUND OF THE INVENTION

Overexpression of Pim kinases is detected in a wide variety ofhematologic and solid cancers. Overexpression of various family membershave been noted in multiple myeloma, AML, pancreatic and hepatocellularcancers. Claudio et al., Blood 2002, 100, 2175-86; Amson et al., Proc.Nat. Acad. Sci. USA, 1989, 86, 8857-61; Mizuki et al., Blood, 2003, 101,3164-73; Li et al., Canc. Res., 2006, 66, 6741-7; Fujii et al., Int. J.Canc., 2005, 114, 209-18. Pim1 overexpression is associated with poorprognosis in mantle cell lymphoma, esophageal and head and neck cancers.Hsi et al., Leuk. Lymph., 2008, 49, 2081-90; Liu et al., J. Surg.Oncol., 2010, 102, 683-88; Peltola et al., Neoplasia, 2009, 11, 629-36.Pim2 overexpression is associated with an aggressive clinical course ina subset of DLBCL patients. Gomez-Abad et al., Blood, 2011, 118,5517-27. Overexpression is often seen where Myc is overexpressed and Pimkinases can convey resistance to traditional chemotherapeutic agents andradiation. Chen et al., Blood, 2009, 114, 4150-57; Isaac et al., DrugResis. Updates, 2011, 14, 203-11; Hsu et al., Cancer Lett., 2012, 319,214; Peltola et al., Neoplasia, 2009, 11, 629-36. As such, these dataindicate that inhibition of Pim kinases will be useful to providetherapeutic benefit in cancer patients.

Pim kinase inhibitors have been described in, for example, US Pat. Pub.Nos. 2014/0200216, 2014/0200227, and 2015/0057265. In particular, thePim-inhibiting compoundN-{4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(including both 7R and 7S diastereomers) is described in US Pat. Pub.No. 2014/0200227. Accordingly, new forms of Pim-inhibiting molecules areneeded to help prepare pharmaceutically useful formulations and dosageforms with suitable properties related to, for example, facilitating themanufacture of safe, effective, high quality drug products. The presentinvention described herein is directed toward this end.

SUMMARY OF THE INVENTION

The present invention is directed to salts ofN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide.

The present invention is further directed to the phosphoric acid salt ofN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide.

The present invention is further directed to crystalline forms of thephosphoric acid salt ofN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide.

The present invention is further directed to the dihydrochloric acidsalt ofN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide.

The present invention is further directed to the monohydrochloric acidsalt ofN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide.

The present invention is further directed to the maleic acid salt,adipic acid salt, hydrobromic acid salt, (R)-(−)-mandelic acid salt,salicylic acid salt, benzoic acid salt, pyroglutamic acid salt,methanesulfonic acid salt, (1S)-(+)-10-camphorsulfonic acid salt,fumaric acid salt, sulfuric acid salt, L-tartaric acid salt, andD-tartaric acid salt ofN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide.

The present invention is further directed to crystalline forms of thesalts described herein.

The present invention is further directed to pharmaceutical compositionscomprising a salt or crystalline form described herein, and at least onepharmaceutically acceptable carrier.

The present invention is further directed to therapeutic methods ofusing the salts and crystalline forms described herein.

The present invention is further directed to processes for preparing thesalts and crystalline forms described herein.

The present invention is further directed to intermediates useful in thepreparation of the salts and crystalline forms described herein.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the XRPD pattern of Compound 1 phosphoric acid salt Form I.

FIG. 2 shows the DSC thermogram of Compound 1 phosphoric acid salt FormI.

FIG. 3 shows the TGA thermogram of Compound 1 phosphoric acid salt FormI.

FIG. 4 shows the XRPD pattern of a solid form of Compound 1dihydrochloric acid salt.

FIG. 5 shows the DSC thermogram of a solid form of Compound 1dihydrochloric acid salt.

FIG. 6 shows the TGA thermogram of a solid form of Compound 1dihydrochloric acid salt.

FIG. 7 shows the XRPD pattern of Compound 1 phosphoric acid salt FormII.

FIG. 8 shows the DSC thermogram of Compound 1 phosphoric acid salt FormII.

FIG. 9 shows the TGA thermogram of Compound 1 phosphoric acid salt FormII.

FIG. 10 shows the XRPD pattern of Compound 1 phosphoric acid salt FormIII.

FIG. 11 shows the DSC thermogram of Compound 1 phosphoric acid salt FormIII.

FIG. 12 shows the TGA thermogram of Compound 1 phosphoric acid salt FormIII.

FIG. 13 shows the XRPD pattern of Compound 1 phosphoric acid salt FormIV.

FIG. 14 shows the DSC thermogram of Compound 1 phosphoric acid salt FormIV.

FIG. 15 shows the TGA thermogram of Compound 1 phosphoric acid salt FormIV.

FIG. 16 shows the XRPD pattern of Compound 1 phosphoric acid salt FormV.

FIG. 17 shows the DSC thermogram of Compound 1 phosphoric acid salt FormV.

FIG. 18 shows the TGA thermogram of Compound 1 phosphoric acid salt FormV.

FIG. 19 shows the XRPD pattern of Compound 1 phosphoric acid salt FormVI.

FIG. 20 shows the DSC thermogram of Compound 1 phosphoric acid salt FormVI.

FIG. 21 shows the TGA thermogram of Compound 1 phosphoric acid salt FormVI.

FIG. 22 shows the XRPD pattern of a solid form of Compound 1mono-hydrochloric acid salt.

FIG. 23 shows the DSC thermogram of a solid form of Compound 1mono-hydrochloric acid salt.

FIG. 24 shows the XRPD pattern of a solid form of Compound 1 maleic acidsalt.

FIG. 25 shows the DSC thermogram of a solid form of Compound 1 maleicacid salt.

FIG. 26 shows the TGA thermogram of a solid form of Compound 1 maleicacid salt.

FIG. 27 shows the XRPD pattern of a solid form of Compound 1 adipic acidsalt.

FIG. 28 shows the DSC thermogram of a solid form of Compound 1 adipicacid salt.

FIG. 29 shows the TGA thermogram of a solid form of Compound 1 adipicacid salt.

FIG. 30 shows the XRPD pattern of a solid form of Compound 1 hydrobromicacid salt.

FIG. 31 shows the DSC thermogram of a solid form of Compound 1hydrobromic acid salt.

FIG. 32 shows the TGA thermogram of a solid form of Compound 1hydrobromic acid salt.

FIG. 33 shows the XRPD pattern of a solid form of Compound 1R-(−)-mandelic acid salt.

FIG. 34 shows the DSC thermogram of a solid form of Compound 1R-(−)-mandelic acid salt.

FIG. 35 shows the TGA thermogram of a solid form of Compound 1R-(−)-mandelic acid salt.

FIG. 36 shows the XRPD pattern of a solid form of Compound 1 salicylicacid salt.

FIG. 37 shows the DSC thermogram of a solid form of Compound 1 salicylicacid salt.

FIG. 38 shows the XRPD pattern of a solid form of Compound 1 benzoicacid salt.

FIG. 39 shows the XRPD pattern of a solid form of Compound 1benzenesulfonic acid salt.

FIG. 40 shows the XRPD pattern of a solid form of Compound 1L-pyroglutamic acid salt.

FIG. 41 shows the XRPD pattern of a solid form of Compound 1methanesulfonic acid salt.

FIG. 42 shows the XRPD pattern of a solid form of Compound 1(1S)-(+)-10-camphorsulfonic acid salt.

FIG. 43 shows the XRPD pattern of a solid form of Compound 1 fumaricacid salt.

FIG. 44 shows the XRPD pattern of a solid form of Compound 1 sulfuricacid salt.

FIG. 45 shows the XRPD pattern of a solid form of Compound 1 L-tartaricacid salt.

FIG. 46 shows the XRPD pattern of a solid form of Compound 1 D-tartaricacid salt.

DETAILED DESCRIPTION

The present invention is directed to, inter alia, salts ofN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1), the structure of which is shown below.

Compound 1 and its salts are a Pim kinase inhibitors useful in thetreatment of diseases in which, for example, one or more Pim kinases(e.g., Pim 1, Pim2, and/or Pim 3) is upregulated. In some embodiments,the salt of Compound 1 provided herein is a solid form. In someembodiments, the present invention relates to a solid form comprising asalt provided herein. In some embodiments, the solid form iscrystalline.

In some embodiments, the salt of the invention is a phosphoric acid saltof Compound 1, such as a monophosphoric acid salt form. Themonophosphoric acid salt form of Compound 1 is referred to herein as“Compound 1 phosphoric acid salt,” “Compound 1 phosphoric acid,” or“Compound 1 phosphate.” An alternative name for the salt isN—{(R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropicolinamidephosphate.

In some embodiments, the salt of the invention is a hydrochloric acidsalt of Compound 1, such as a dihydrochloric acid salt form. Thedihydrochloric acid salt form of Compound 1 is referred to herein as“Compound 1 dihydrochloric acid salt,” “Compound 1 dihydrochloric acid,”or “Compound 1 dihydrochloride.” An alternative name for the salt isN—{(R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropicolinamidedihydrochloride.

In some embodiments, the hydrochloric acid salt of Compound 1 is amonohydrochloric acid salt of Compound 1. The monohydrochloric acid saltform of Compound 1 is referred to herein as “Compound 1 monohydrochloricacid salt,” “Compound 1 monohydrochloric acid,” or “Compound 1monohydrochloride.” An alternative name for the salt isN—{(R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropicolinamidemono-hydrochloride.

In some embodiments, the salt of the invention is a maleic(cis-butenedioic) acid salt of Compound 1. The maleic acid salt form ofCompound 1 is referred to herein as “Compound 1 maleic acid salt,”“Compound 1 maleic acid,” or “Compound 1 maleate.” An alternative namefor the salt isN—{(R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropicolinamidemaleate.

In some embodiments, the salt of the invention is an adipic acid salt ofCompound 1. The adipic acid salt form of Compound 1 is referred toherein as “Compound 1 adipic acid salt,” “Compound 1 adipic acid,” or“Compound 1 adipate.” An alternative name for the salt isN—{(R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropicolinamideadipate.

In some embodiments, the salt of the invention is a hydrobromic acidsalt of Compound 1. In some embodiments, the hydrobromic acid salt ofCompound 1 a dihydrobromic acid salt form. In some embodiments, thehydrobromic acid salt of Compound 1 a monohydrobromic acid salt form.The hydrobromic acid salt form of Compound 1 is referred to herein as“Compound 1 hydrobromic acid salt,” “Compound 1 hydrobromic acid,” or“Compound 1 hydrobromide.” An alternative name for the salt isN—{(R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropicolinamidehydrobromide.

In some embodiments, the salt of the invention is a (R)-(−)-mandelicacid salt of Compound 1. In some embodiments, the (R)-(−)-mandelic acidsalt of Compound 1 is a mono-(R)-(−)-mandelic acid salt. In someembodiments, the (R)-(−)-mandelic acid salt of Compound 1 is adi-(R)-(−)-mandelic acid salt. The (R)-(−)-mandelic acid salt form ofCompound 1 is referred to herein as “Compound 1 mandelic acid salt,”“Compound 1 mandelic acid,” or “Compound 1 mandelate.” An alternativename for the salt isN—{(R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropicolinamidemandelate.

In some embodiments, the salt of the invention is a salicylic acid saltof Compound 1. In some embodiments, the salicylic acid salt of Compound1 is a mono-salicylic acid salt. In some embodiments, the salicylic acidsalt of Compound 1 is a di-salicylic acid salt. The salicylic acid saltform of Compound 1 is referred to herein as “Compound 1 salicylic acidsalt,” “Compound 1 salicylic acid,” or “Compound 1 salicylate.” Analternative name for the salt isN—{(R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropicolinamidesalicylate.

In some embodiments, the salt of the invention is a benzoic acid salt ofCompound 1. In some embodiments, the benzoic acid salt of Compound 1 isa mono-benzoic acid salt. In some embodiments, the benzoic acid salt ofCompound 1 is a di-benzoic acid salt. The benzoic acid salt form ofCompound 1 is referred to herein as “Compound 1 benzoic acid salt,”“Compound 1 benzoic acid,” or “Compound 1 benzoate.” An alternative namefor the salt isN—{(R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropicolinamidebenzoate.

In some embodiments, the salt of the invention is a benzenesulfonic acidsalt of Compound 1. In some embodiments, the benzenesulfonic acid saltof Compound 1 is a mono-benzenesulfonic acid salt. In some embodiments,the benzenesulfonic acid salt of Compound 1 is a di-benzenesulfonic acidsalt. The benzenesulfonic acid salt form of Compound 1 is referred toherein as “Compound 1 benzenesulfonic acid salt,” “Compound 1benzenesulfonic acid,” or “Compound 1 besylate.” An alternative name forthe salt isN—{(R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropicolinamidebesylate.

In some embodiments, the salt of the invention is L-pyroglutamic acidsalt of Compound 1. In some embodiments, the L-pyroglutamic acid salt ofCompound 1 is a mono-L-pyroglutamic acid salt. In some embodiments, theL-pyroglutamic acid salt of Compound 1 is a di-L-pyroglutamic acid salt.The L-pyroglutamic acid salt form of Compound 1 is referred to herein as“Compound 1 L-pyroglutamic acid salt,” “Compound 1 L-pyroglutamic acid,”or “Compound 1 L-pyroglutamate.” An alternative name for the salt isN—{(R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropicolinamideL-pyroglutamate.

In some embodiments, the salt of the invention is a methanesulfonic acidsalt of Compound 1. In some embodiments, the methanesulfonic acid saltof Compound 1 is a mono-methanesulfonic acid salt form. In someembodiments, the methanesulfonic acid salt of Compound 1 is adi-methanesulfonic acid salt form. The methanesulfonic acid salt form ofCompound 1 is referred to herein as “Compound 1 methanesulfonic acidsalt,” “Compound 1 methanesulfonic acid,” or “Compound 1 mesylate.” Analternative name for the salt isN—{(R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropicolinamidemesylate.

In some embodiments, the salt of the invention is a (1S)-(+)-10-camphorsulfonic acid salt of Compound 1. In some embodiments,the (1S)-(+)-10-camphorsulfonic acid salt of Compound 1 is amono-(1S)-(+)-10-camphorsulfonic acid salt. In some embodiments, the(1S)-(+)-10-camphorsulfonic acid salt of Compound 1 is a(1S)-(+)-10-di-camphorsulfonic acid salt. The(1S)-(+)-10-camphorsulfonic acid salt form of Compound 1 is referred toherein as “Compound 1 (1S)-(+)-10-camphorsulfonic acid salt,” “Compound1 (1S)-(+)-10-camphorsulfonic acid,” or “Compound 1 camsylate.” Analternative name for the salt isN—{(R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropicolinamidecamsylate.

In some embodiments, the salt of the invention is a fumaric(trans-butenedioic) acid salt of Compound 1. The fumaric acid salt formof Compound 1 is referred to herein as “Compound 1 fumaric acid salt,”“Compound 1 fumaric acid,” or “Compound 1 fumarate.” An alternative namefor the salt isN—{(R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropicolinamidefumarate.

In some embodiments, the salt of the invention is a sulfuric acid saltof Compound 1. In some embodiments, the sulfuric acid salt of Compound 1is a mono-sulfuric acid salt form. The sulfuric acid salt form ofCompound 1 is referred to herein as “Compound 1 sulfuric acid salt,”“Compound 1 sulfuric acid,” or “Compound 1 sulfate.” An alternative namefor the salt isN—{(R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropicolinamidesulfate.

In some embodiments, the salt of the invention is a tartaric acid saltof Compound 1, such as L-tartaric acid salt form. The L-tartaric acidsalt form of Compound 1 is referred to herein as “Compound 1 L-tartaricacid salt,” “Compound 1 L-tartaric acid,” or “Compound 1 L-tartrate.” Analternative name for the salt isN—{(R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropicolinamideL-tartrate.

In some embodiments, the tartaric acid salt of Compound 1 is D-tartaricacid salt of Compound 1. The D-tartaric acid salt form of Compound 1 isreferred to herein as “Compound 1 D-tartaric acid salt,” “Compound 1D-tartaric acid,” or “Compound 1 D-tartrate.” An alternative name forthe salt isN—{(R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropicolinamideD-tartrate.

The salts of the invention can be isolated as one or more solid forms.As used herein, the phrase “solid form” refers to a salt of theinvention in either an amorphous state or a crystalline state(“crystalline form” or “crystalline solid”), whereby a salt of theinvention in a crystalline state may optionally include solvent or waterwithin the crystalline lattice, for example, to form a solvated orhydrated crystalline form. The term “hydrated,” as used herein, is meantto refer to a crystalline form that includes water molecules in thecrystalline lattice. Example “hydrated” crystalline forms includehemihydrates, monohydrates, dihydrates, and the like. Other hydratedforms such as channel hydrates and the like are also included within themeaning of the term.

The different crystalline forms of the salts of the invention arecharacterized by X-ray powder diffraction (XRPD), differential scanningcalorimetry (DSC), and/or thermogravimetric analysis (TGA). An X-raypowder diffraction (XRPD) pattern of reflections (peaks) is typicallyconsidered a fingerprint of a particular crystalline form. It is wellknown that the relative intensities of the XRPD peaks can widely varydepending on, inter alia, the sample preparation technique, crystal sizedistribution, various filters used, the sample mounting procedure, andthe particular instrument employed. In some instances, new peaks may beobserved or existing peaks may disappear depending on the type ofinstrument or the settings (for example, whether a Ni filter is used ornot). As used herein, the term “peak” or “characteristic peak” refers toa reflection having a relative height/intensity of at least about 3% ofthe maximum peak height/intensity. Moreover, instrument variation andother factors can affect the 2-theta values. Thus, peak assignments,such as those reported herein, can vary by plus or minus about 0.2°(2-theta), and the term “substantially” or “about” as used in thecontext of XRPD herein is meant to refer to the above-mentionedvariations.

In the same way, temperature readings in connection with DSC, TGA, orother thermal experiments can vary about ±3° C. depending on theinstrument, particular settings, sample preparation, etc. Accordingly, acrystalline form reported herein having a DSC thermogram “substantially”as shown in any of the Figures is understood to accommodate suchvariation.

The salts and compounds disclosed herein can include all isotopes ofatoms occurring within them. Isotopes include those atoms having thesame atomic number but different mass numbers. For example, isotopes ofhydrogen include tritium and deuterium. Salts and compounds of theinvention can also include all isotopes of atoms occurring in theintermediates or final compounds. Isotopes include those atoms havingthe same atomic number but different mass numbers. For example, isotopesof hydrogen include tritium and deuterium. One or more constituent atomsof the compounds of the invention can be replaced or substituted withisotopes of the atoms in natural or non-natural abundance. In someembodiments, the compound includes at least one deuterium atom. Forexample, one or more hydrogen atoms in a compound of the presentdisclosure can be replaced or substituted by deuterium. In someembodiments, the compound includes two or more deuterium atoms. In someembodiments, the compound includes 1, 2, 3, 4, 5, 6, 7 or 8 deuteriumatoms. Synthetic methods for including isotopes into organic compoundsare known in the art.

In some embodiments, the compounds of the invention, or salts thereof,are substantially isolated. By “substantially isolated” is meant thatthe compound or salt is at least partially or substantially separatedfrom the environment in which it was formed or detected. Partialseparation can include, e.g., a composition enriched in the compounds ofthe invention. Substantial separation can include compositionscontaining at least about 50%, at least about 60%, at least about 70%,at least about 80%, at least about 90%, at least about 95%, at leastabout 97%, or at least about 99% by weight of the compounds or salts ofthe invention.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

As used herein, the term “melting point” refers to an endothermic eventor endothermal event observed in e.g., a DSC experiment. An endothermicevent is a process or reaction in which a sample absorbs energy from itssurrounding in the form of e.g., heat as in a DSC experiment. Anexothermic event is a process or reaction in which a sample releasesenergy. The process of heat absorption and release can be detected byDSC. In some embodiments, the term “melting point” is used to describethe major endothermic event revealed on a particular DSC thermogram.

The term “room temperature” as used herein, is understood in the art,and refers generally to a temperature, e.g., a reaction temperature,that is about the temperature of the room in which the reaction iscarried out, for example, a temperature from about 20° C. to about 30°C. The term “elevated temperature” as used herein, is understood in theart, and refer generally to a temperature, e.g., a reaction temperature,that is above room temperature, e.g., above 30° C.

Phosphoric acid salt The present invention is directed to, inter alia, aphosphoric acid salt ofN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide,such as the salt which is shown below.

Compound 1 phosphoric acid salt can be prepared as an amorphous solid,as a crystalline solid, or as a mixture thereof. In some embodiments,the crystalline solid has Form I, which is described below in theExamples. In some embodiments, the crystalline solid having Form I has acharacteristic XRPD peak, in terms of 2-theta, at about 4.6 degrees. Insome embodiments, the crystalline solid having Form I has at least onecharacteristic XRPD peak, in terms of 2-theta, at about 4.6 or about 9.4degrees. In some embodiments, the crystalline solid having Form I has atleast one characteristic XRPD peak, in terms of 2-theta, at about 4.6,about 9.4, or about 13.1 degrees. In some embodiments, the crystallinesolid of Form I has two or more characteristic XRPD peaks, in terms of2-theta, selected from about 4.6, about 9.4, about 13.1, about 16.2,about 17.4, about 17.9, about 18.8, about 19.4, about 21.1, about 23.0,and about 24.8. In some embodiments, the crystalline solid of Form I hasthree or more characteristic XRPD peaks, in terms of 2-theta, selectedfrom about 4.6, about 9.4, about 13.1, about 16.2, about 17.4, about17.9, about 18.8, about 19.4, about 21.1, about 23.0, about 24.8, andabout 25.2 degrees. In some embodiments, the crystalline solid of Form Ihas four or more characteristic XRPD peaks, in terms of 2-theta,selected from about 4.6, about 9.4, about 13.1, about 16.2, about 17.4,about 17.9, about 18.8, about 19.4, about 21.1, about 23.0, about 24.8,about 25.2 degrees. In some embodiments, the crystalline solid of Form Ihas an XRPD pattern substantially as depicted in FIG. 1. In someembodiments, the crystalline solid of Form I has a melting point ofabout 250° C. In some embodiments, the crystalline solid of Form I hasan endothermic event at about 198° C. or about 250° C. In someembodiments, the crystalline solid of Form I has a DSC thermogramsubstantially as depicted in FIG. 2. In some embodiments, thecrystalline solid of Form I has a TGA thermogram substantially asdepicted in FIG. 3.

The advantages of the phosphoric acid salt include high crystallinity,high melting point, stable crystalline form (e.g., Form I), andnon-hygroscopic properties, each of which facilitate the purification,reproducibility, scale up, manufacturing, and formulation of the drugcompound.

In some embodiments, Compound 1 phosphoric acid salt is a crystallinesolid having Form II. In some embodiments, the crystalline solid havingForm II has at least one characteristic XRPD peak, in terms of 2-theta,selected from about 4.7, about 9.4, about 18.8, about 21.7, about 24.8,and about 33.3 degrees. In some embodiments, the crystalline solidhaving Form II has at least 2 characteristic XRPD peaks, in terms of2-theta, selected from about 4.7, about 9.4, about 18.8, about 21.7,about 24.8, and about 33.3 degrees. In some embodiments, the crystallinesolid having Form II has at least 3 characteristic XRPD peaks, in termsof 2-theta, selected from about 4.7, about 9.4, about 18.8, about 21.7,about 24.8, and about 33.3 degrees. In some embodiments, the crystallinesolid having Form II has at least one characteristic XRPD peak, in termsof 2-theta, selected from about 4.7, about 9.4, about 18.8 degrees. Insome embodiments, the crystalline solid having Form II has at least twocharacteristic XRPD peaks, in terms of 2-theta, selected from about 4.7,about 9.4, about 18.8 degrees. In some embodiments, the crystallinesolid having Form II has an XRPD pattern as depicted in FIG. 7. In someembodiments, the crystalline solid having Form II has a melting point ofabout 249° C. In some embodiments, the crystalline solid having Form IIhas an endothermic event at about 249° C. In some embodiments, thecrystalline solid having Form II has a DSC thermogram substantially asdepicted in FIG. 8. In some embodiments, the crystalline solid havingForm II has a TGA thermogram substantially as depicted in FIG. 9.

In some embodiments, Compound 1 phosphoric acid salt is a crystallinesolid having Form III. In some embodiments, the crystalline solid havingForm III has at least one characteristic XRPD peak, in terms of 2-theta,selected from about 4.6, about 9.4, about 13.3, about 16.3, about 18.9,about 19.2, about 21.2, about 22.5, about 23.1, about 24.9, and about26.7 degrees. In some embodiments, the crystalline solid having Form IIIhas at least two characteristic XRPD peaks, in terms of 2-theta,selected from about 4.6, about 9.4, about 13.3, about 16.3, about 18.9,about 19.2, about 21.2, about 22.5, about 23.1, about 24.9, and about26.7 degrees. In some embodiments, the crystalline solid having Form IIIhas at least three characteristic XRPD peaks, in terms of 2-theta,selected from about 4.6, about 9.4, about 13.3, about 16.3, about 18.9,about 19.2, about 21.2, about 22.5, about 23.1, about 24.9, and about26.7 degrees. In some embodiments, the crystalline solid having Form IIIhas at least one characteristic XRPD peak, in terms of 2-theta, selectedfrom about 4.6, about 18.9, about 19.2, about 22.5, and about 23.1degrees. In some embodiments, the crystalline solid having Form III hasat least two characteristic XRPD peaks, in terms of 2-theta, selectedfrom about 4.6, about 18.9, about 19.2, about 22.5, and about 23.1degrees. In some embodiments, the crystalline solid having Form III hasat least three characteristic XRPD peaks, in terms of 2-theta, selectedfrom about 4.6, about 18.9, about 19.2, about 22.5, and about 23.1degrees. In some embodiments, the crystalline solid having Form III hasan XRPD pattern as depicted in FIG. 10. In some embodiments, thecrystalline solid having Form III has a melting point of about 250° C.In some embodiments, the crystalline solid having Form III has anendothermic event at about 250° C. In some embodiments, the crystallinesolid having Form III has a DSC thermogram substantially as depicted inFIG. 11. In some embodiments, the crystalline solid having Form III hasa TGA thermogram substantially as depicted in FIG. 12.

In some embodiments, Compound 1 phosphoric acid salt is a crystallinesolid having Form IV. In some embodiments, the crystalline solid havingForm IV has at least one characteristic XRPD peak, in terms of 2-theta,selected from about 4.1, about 13.3, about 16.4, about 17.7, about 18.6,about 19.8, about 21.4, and about 23.3 degrees. In some embodiments, thecrystalline solid having Form IV has at least two characteristic XRPDpeaks, in terms of 2-theta, selected from about 4.1, about 13.3, about16.4, about 17.7, about 18.6, about 19.8, about 21.4, and about 23.3degrees. In some embodiments, the crystalline solid having Form IV hasat least three characteristic XRPD peaks, in terms of 2-theta, selectedfrom about 4.1, about 13.3, about 16.4, about 17.7, about 18.6, about19.8, about 21.4, and about 23.3 degrees. In some embodiments, thecrystalline solid having Form IV has at least one characteristic XRPDpeak, in terms of 2-theta, selected from about 4.1, about 13.3, about16.4, about 18.6, about 19.8, and about 21.4 degrees. In someembodiments, the crystalline solid having Form IV has at least twocharacteristic XRPD peaks, in terms of 2-theta, selected from about 4.1,about 13.3, about 16.4, about 18.6, about 19.8, and about 21.4 degrees.In some embodiments, the crystalline solid having Form IV has at leastthree characteristic XRPD peaks, in terms of 2-theta, selected fromabout 4.1, about 13.3, about 16.4, about 18.6, about 19.8, and about21.4 degrees. In some embodiments, the crystalline solid having Form IVhas an XRPD pattern as depicted in FIG. 13. In some embodiments, thecrystalline solid having Form IV has a melting point of about 245° C. Insome embodiments, the crystalline solid having Form IV has a meltingpoint of about 245° C. In some embodiments, the crystalline solid havingForm IV has a DSC thermogram substantially as depicted in FIG. 14. Insome embodiments, the crystalline solid having Form IV has a TGAthermogram substantially as depicted in FIG. 15.

In some embodiments, Compound 1 phosphoric acid salt is a crystallinesolid having Form V. In some embodiments, the crystalline solid havingForm V has at least one characteristic XRPD peak, in terms of 2-theta,selected from about 7.3, about 10.9, about 16.4, about 18.5, about 19.8,about 22.6, and about 26.1 degrees. In some embodiments, the crystallinesolid having Form V has at least two characteristic XRPD peaks, in termsof 2-theta, selected from about 7.3, about 10.9, about 16.4, about 18.5,about 19.8, about 22.6, and about 26.1 degrees. In some embodiments, thecrystalline solid having Form V has at least three characteristic XRPDpeaks, in terms of 2-theta, selected from about 7.3, about 10.9, about16.4, about 18.5, about 19.8, about 22.6, and about 26.1 degrees. Insome embodiments, the crystalline solid having Form V has at least onecharacteristic XRPD peak, in terms of 2-theta, selected from about 7.3,about 10.9, about 16.4, about 18.5, and about 19.8 degrees. In someembodiments, the crystalline solid having Form V has at least twocharacteristic XRPD peaks, in terms of 2-theta, selected from about 7.3,about 10.9, about 16.4, about 18.5, and about 19.8 degrees. In someembodiments, the crystalline solid having Form V has at least threecharacteristic XRPD peaks, in terms of 2-theta, selected from about 7.3,about 10.9, about 16.4, about 18.5, and about 19.8 degrees. In someembodiments, the crystalline solid having Form V has an XRPD pattern asshown on FIG. 16. In some embodiments, the crystalline solid having FormV has an endothermic event at about 95° C. or about 245° C. In someembodiments, the crystalline solid having Form V has a DSC thermogramsubstantially as depicted in FIG. 17. In some embodiments, thecrystalline solid having Form V has a TGA thermogram substantially asdepicted in FIG. 18.

In some embodiments, Compound 1 phosphoric acid salt is a crystallinesolid having Form VI. In some embodiments, the crystalline solid havingForm VI has at least one characteristic XRPD peak, in terms of 2-theta,selected from about 6.5, about 8.3, about 10.7, about 13.2, about 17.3,and about 19.1 degrees. In some embodiments, the crystalline solidhaving Form VI has at least two characteristic XRPD peaks, in terms of2-theta, selected from about 6.5, about 8.3, about 10.7, about 13.2,about 17.3, and about 19.1 degrees. In some embodiments, the crystallinesolid having Form VI has at least three characteristic XRPD peaks, interms of 2-theta, selected from about 6.5, about 8.3, about 10.7, about13.2, about 17.3, and about 19.1 degrees. In some embodiments, thecrystalline solid having Form VI has at least one characteristic XRPDpeak, in terms of 2-theta, selected from about 6.5, about 8.3, and about10.7 degrees. In some embodiments, the crystalline solid having Form VIhas at least two characteristic XRPD peaks, in terms of 2-theta,selected from about 6.5, about 8.3, and about 10.7 degrees. In someembodiments, the crystalline solid having Form VI has an XRPD pattern asshown on FIG. 19. In some embodiments, the crystalline solid having FormVI has a melting point of about 86° C. In some embodiments, thecrystalline solid having Form VI has an endothermic event at about 86°C. or about 221° C. In some embodiments, the crystalline solid havingForm VI has a DSC thermogram substantially as depicted in FIG. 20. Insome embodiments, the crystalline solid having Form VI has a TGAthermogram substantially as depicted in FIG. 21.

In some embodiment, the present invention provides a mixture ofcrystalline solid Form I and one or more solid forms selected fromamorphous, Form II, Form III, Form IV, and Form V.

In some embodiments, the mixture of the crystalline solid Form I hasgreater than about 50%, about 60%, about 70%, about 80%, about 90%,about 95%, about 98%, or about 99% of Form I.

In some embodiment, crystalline solid Form I is prepared in high purity.Purity values indicate the percentage of the amount of sample that isForm I. Purity values can be determined, for example, by HPLC/UVmethods. In some embodiments, Form I has a purity greater than about90%, greater than about 95%, greater than about 97%, greater than about98%, or greater than about 99%. In some embodiments, Form I issubstantially free of impurities, such as organic impurities, inorganicimpurities, and/or residual solvents. Examples of organic impuritiesinclude e.g., starting materials and process intermediates such as

Examples of organic impurities include e.g., process impurities such as

Examples of inorganic impurities include e.g., heavy metals, palladium,and ruthenium. Examples of residual solvents include e.g., acetonitrile,dichloromethane, N,N-dimethylformamide, 1,4-dioxane, n-heptane,methanol, and 2-propanol.

Hydrochloric Acid Salts

The present invention further provides the dihydrochloric acid salt ofCompound 1. In some embodiments, a solid form of the dihydrochloric acidsalt has an XRPD pattern as shown on FIG. 4. In some embodiments, asolid form of the dihydrochloric acid salt has a melting point of about213° C. In some embodiments, a solid form of the dihydrochloric acidsalt has an endothermic event at about 213° C. In some embodiments, asolid form of the dihydrochloric acid salt is characterized by a DSCthermogram substantially as depicted in FIG. 5. In some embodiments, asolid form of the dihydrochloric acid salt is characterized by a TGAthermogram substantially as depicted in FIG. 6.

In some embodiments, the dihydrochloric acid salt of Compound 1 is acrystalline solid. In some embodiments, the crystalline solid ofCompound 1 dihydrochloric acid salt has at least one characteristic XRPDpeak, in terms of 2-theta, selected from about 8.3, about 18.9, andabout 25.0 degrees. In some embodiments, the crystalline solid ofCompound 1 dihydrochloric acid salt has two or more characteristic XRPDpeaks, in terms of 2-theta, selected from about 8.3, about 18.9, andabout 25.0 degrees.

The present invention further provides the monohydrochloric acid salt ofCompound 1. In some embodiments, a solid form of the monohydrochloricacid salt has an XRPD pattern as shown on FIG. 22. In some embodiments,a solid form of the monohydrochloric acid salt has a melting point ofabout 209° C. In some embodiments, a solid form of the monohydrochloricacid salt has an endothermic event at about 209° C. In some embodiments,a solid form of the monohydrochloric acid salt is characterized by a DSCthermogram substantially as depicted in FIG. 23.

In some embodiments, the monohydrochloric acid salt of Compound 1 is acrystalline solid. In some embodiments, the crystalline solid ofCompound 1 monohydrochloric acid salt has at least one characteristicXRPD peak, in terms of 2-theta, selected from about 7.8, about 8.8,about 12.6, about 14.5, about 17.4, about 23.8, and about 25.2 degrees.In some embodiments, the crystalline solid of Compound 1monohydrochloric acid salt has two or more characteristic XRPD peaks, interms of 2-theta, selected from about 7.8, about 8.8, about 12.6, about14.5, about 17.4, about 23.8, and about 25.2 degrees. In someembodiments, the crystalline solid of Compound 1 monohydrochloric acidsalt has three or more characteristic XRPD peaks, in terms of 2-theta,selected from about 7.8, about 8.8, about 12.6, about 14.5, about 17.4,about 23.8, and about 25.2 degrees.

Maleic Acid Salt

The present invention further provides a maleic acid salt of Compound 1.In some embodiments, a solid form of the maleic acid salt has an XRPDpattern as shown on FIG. 24. In some embodiments, a solid form of themaleic acid salt has a melting point of about 202° C. In someembodiments, a solid form of the maleic acid salt has an endothermicevent at about 202° C. In some embodiments, a solid form of the maleicacid salt has a DSC thermogram substantially as depicted in FIG. 25. Insome embodiments, the maleic acid salt has a TGA thermogramsubstantially as depicted in FIG. 26.

In some embodiments, the maleic acid salt of Compound 1 is a crystallinesolid. In some embodiments, the crystalline solid of Compound 1 maleicacid salt has at least one characteristic XRPD peak, in terms of2-theta, selected from about 9.0, about 9.5, about 11.2, about 14.8,about 15.9, about 18.5, about 19.5, about 19.9, about 21.3, about 22.9,about 24.8, about 25.8, about 27.6, and about 30.9 degrees. In someembodiments, the crystalline solid of Compound 1 maleic acid salt has atleast two characteristic XRPD peaks, in terms of 2-theta, selected fromabout 9.0, about 9.5, about 11.2, about 14.8, about 15.9, about 18.5,about 19.5, about 19.9, about 21.3, about 22.9, about 24.8, about 25.8,about 27.6, and about 30.9 degrees. In some embodiments, the crystallinesolid of Compound 1 maleic acid salt has at least three characteristicXRPD peaks, in terms of 2-theta, selected from about 9.0, about 9.5,about 11.2, about 14.8, about 15.9, about 18.5, about 19.5, about 19.9,about 21.3, about 22.9, about 24.8, about 25.8, about 27.6, and about30.9 degrees.

Adipic Acid Salt

The present invention further provides an adipic acid salt ofCompound 1. In some embodiments, a solid form of the adipic acid salthas an XRPD pattern as shown on FIG. 27. In some embodiments, a solidform of the adipic acid salt has a melting point of about 182° C. Insome embodiments, a solid form of the adipic acid salt has anendothermic event at about 150° C. or about 182° C. In some embodiments,a solid form of the adipic acid salt has a DSC thermogram substantiallyas depicted in FIG. 28. In some embodiments, the adipic acid salt has aTGA thermogram substantially as depicted in FIG. 29.

In some embodiments, the adipic acid salt of Compound 1 is a crystallinesolid. In some embodiments, the crystalline solid of Compound 1 adipicacid salt has at least one characteristic XRPD peak, in terms of2-theta, selected from about 9.3, about 15.0, about 16.2, about 17.6,about 18.7, about 20.0, about 22.1, about 22.7, about 24.3, about 24.9,about 27.1, and about 28.7 degrees. In some embodiments, the crystallinesolid of Compound 1 adipic acid salt has at least two characteristicXRPD peaks, in terms of 2-theta, selected from about 9.3, about 15.0,about 16.2, about 17.6, about 18.7, about 20.0, about 22.1, about 22.7,about 24.3, about 24.9, about 27.1, and about 28.7 degrees. In someembodiments, the crystalline solid of Compound 1 adipic acid salt has atleast three characteristic XRPD peaks, in terms of 2-theta, selectedfrom about 9.3, about 15.0, about 16.2, about 17.6, about 18.7, about20.0, about 22.1, about 22.7, about 24.3, about 24.9, about 27.1, andabout 28.7 degrees.

Hydrobromic Acid Salt The present invention further provides ahydrobromic acid salt of Compound 1. In some embodiments, a solid formof the hydrobromic acid salt has an XRPD pattern as shown on FIG. 32. Insome embodiments, a solid form of the hydrobromic acid salt has amelting point of about 247° C. In some embodiments, a solid form of thehydrobromic acid salt has a DSC thermogram substantially as depicted inFIG. 33. In some embodiments, a solid form of the hydrobromic acid salthas a TGA thermogram substantially as depicted in FIG. 34.

In some embodiments, the hydrobromic acid salt of Compound 1 is acrystalline solid. In some embodiments, the crystalline solid ofCompound 1 hydrobromic acid salt has at least one characteristic XRPDpeak, in terms of 2-theta, selected from about 6.5, about 9.5, about12.9, about 16.6, about 17.9, about 19.5, about 21.7, about 22.5, about23.7, about 24.3, about 26.5, about 27.5, and about 28.3 degrees. Insome embodiments, the crystalline solid of Compound 1 hydrobromic acidsalt has at least two characteristic XRPD peaks, in terms of 2-theta,selected from about 6.5, about 9.5, about 12.9, about 16.6, about 17.9,about 19.5, about 21.7, about 22.5, about 23.7, about 24.3, about 26.5,about 27.5, and about 28.3 degrees. In some embodiments, the crystallinesolid of Compound 1 hydrobromic acid salt has at least threecharacteristic XRPD peaks, in terms of 2-theta, selected from about 6.5,about 9.5, about 12.9, about 16.6, about 17.9, about 19.5, about 21.7,about 22.5, about 23.7, about 24.3, about 26.5, about 27.5, and about28.3 degrees.

(R)-(−)-Mandelic Acid Salt

The present invention further provides a (R)-(−)-mandelic acid salt ofCompound 1. In some embodiments, a solid form of the (R)-(−)-mandelicacid salt has an XRPD pattern as shown on FIG. 33. In some embodiments,a solid form of the (R)-(−)-mandelic acid salt has a melting point ofabout 224° C. In some embodiments, a solid form of the (R)-(−)-mandelicacid salt has an endothermic event at about 223° C. or about 225° C. Insome embodiments, a solid form of the (R)-(−)-mandelic acid salt has aDSC thermogram substantially as depicted in FIG. 34. In someembodiments, a solid form of the (R)-(−)-mandelic acid salt has a TGAthermogram substantially as depicted in FIG. 35.

In some embodiments, the (R)-(−)-mandelic acid salt of Compound 1 is acrystalline solid. In some embodiments, the crystalline solid ofCompound 1 (R)-(−)-mandelic acid salt has at least one characteristicXRPD peak, in terms of 2-theta, selected from about 11.2, about 13.8,about 18.6, about 20.6, about 22.5, and about 24.1 degrees. In someembodiments, the crystalline solid of Compound 1 (R)-(−)-mandelic acidsalt has at least two characteristic XRPD peaks, in terms of 2-theta,selected from about 11.2, about 13.8, about 18.6, about 20.6, about22.5, and about 24.1 degrees. In some embodiments, the crystalline solidof Compound 1 (R)-(−)-mandelic acid salt has at least threecharacteristic XRPD peaks, in terms of 2-theta, selected from about11.2, about 13.8, about 18.6, about 20.6, about 22.5, and about 24.1degrees.

Salicylic Acid Salt

In some embodiments, the salt of Compound 1 is a salicylic acid salt. Insome embodiments, a solid form of the salicylic acid salt has an XRPDpattern as shown on FIG. 36. In some embodiments, a solid form of thesalicylic acid salt has an endothermic event at about 180° C. or about208° C. In some embodiments, a solid form of the salicylic acid salt hasa DSC thermogram substantially as depicted in FIG. 37.

In some embodiments, the salicylic acid salt of Compound 1 is acrystalline solid. In some embodiments, the crystalline solid ofCompound 1 salicylic acid salt has at least one characteristic XRPDpeak, in terms of 2-theta, selected from about 21.2 and about 23.5degrees. In some embodiments, the crystalline solid of Compound 1salicylic acid salt has at least one characteristic XRPD peak, in termsof 2-theta, selected from about 11.8, about 16.7, about 18.7, about21.2, about 21.9, about 23.0, about 23.5, and about 24.1 degrees. Insome embodiments, the crystalline solid of Compound 1 salicylic acidsalt has at least two characteristic XRPD peaks, in terms of 2-theta,selected from about 11.8, about 16.7, about 18.7, about 21.2, about21.9, about 23.0, about 23.5, and about 24.1 degrees. In someembodiments, the crystalline solid of Compound 1 salicylic acid salt hasat least three characteristic XRPD peaks, in terms of 2-theta, selectedfrom about 11.8, about 16.7, about 18.7, about 21.2, about 21.9, about23.0, about 23.5, and about 24.1 degrees.

Other Salts

The present invention further provides a benzoic acid salt ofCompound 1. In some embodiments, a solid form of the benzoic acid salthas an XRPD pattern as shown on FIG. 38. In some embodiments, thebenzoic acid salt of Compound 1 is a crystalline solid. In someembodiments, the crystalline solid of Compound 1 benzoic acid salt hasat least one characteristic XRPD peak, in terms of 2-theta, selectedfrom about 11.6, about 14.9, about 16.9, about 18.8, about 21.5, about23.2, about 23.7, and about 24.9 degrees. In some embodiments, thecrystalline solid of Compound 1 benzoic acid salt has at least twocharacteristic XRPD peaks, in terms of 2-theta, selected from about11.6, about 14.9, about 16.9, about 18.8, about 21.5, about 23.2, about23.7, and about 24.9 degrees. In some embodiments, the crystalline solidof Compound 1 benzoic acid salt has at least three characteristic XRPDpeaks, in terms of 2-theta, selected from about 11.6, about 14.9, about16.9, about 18.8, about 21.5, about 23.2, about 23.7, and about 24.9degrees.

The present invention further provides a benzenesulfonic acid salt ofCompound 1. In some embodiments, a solid form of the benzenesulfonicacid salt has an XRPD pattern as shown on FIG. 39. In some embodiments,the benzenesulfonic acid salt of Compound 1 is a crystalline solid. Insome embodiments, the crystalline solid of Compound 1 benzenesulfonicacid salt has at least one characteristic XRPD peak, in terms of2-theta, selected from about 6.6, about 9.1, about 12.9, about 13.3,about 14.5, about 18.0, about 23.5, and about 23.9 degrees. In someembodiments, the crystalline solid of Compound 1 benzenesulfonic acidsalt has at least two characteristic XRPD peaks, in terms of 2-theta,selected from about 6.6, about 9.1, about 12.9, about 13.3, about 14.5,about 18.0, about 23.5, and about 23.9 degrees. In some embodiments, thecrystalline solid of Compound 1 benzenesulfonic acid salt has at leastthree characteristic XRPD peaks, in terms of 2-theta, selected fromabout 6.6, about 9.1, about 12.9, about 13.3, about 14.5, about 18.0,about 23.5, and about 23.9 degrees.

The present invention further provides an L-pyroglutamic acid salt ofCompound 1. In some embodiments, a solid form of the L-pyroglutamic acidsalt has an XRPD pattern as shown on FIG. 40. In some embodiments, theL-pyroglutamic acid salt of Compound 1 is a crystalline solid. In someembodiments, the crystalline solid of Compound 1 L-pyroglutamic acidsalt has at least one characteristic XRPD peak, in terms of 2-theta,selected from about 4.4, about 10.7, about 11.5, about 18.0, about 20.7,about 21.2, and about 22.9 degrees. In some embodiments, the crystallinesolid of Compound 1 L-pyroglutamic acid salt has at least twocharacteristic XRPD peaks, in terms of 2-theta, selected from about 4.4,about 10.7, about 11.5, about 18.0, about 20.7, about 21.2, and about22.9 degrees. In some embodiments, the crystalline solid of Compound 1L-pyroglutamic acid salt has at least three characteristic XRPD peaks,in terms of 2-theta, selected from about 4.4, about 10.7, about 11.5,about 18.0, about 20.7, about 21.2, and about 22.9 degrees.

The present invention further provides a methanesulfonic acid salt ofCompound 1. In some embodiments, a solid form of the methanesulfonicacid salt has an XRPD pattern as shown on FIG. 41. In some embodiments,the methanesulfonic acid salt of Compound 1 is a crystalline solid. Insome embodiments, the crystalline solid of Compound 1 methanesulfonicacid salt has at least one characteristic XRPD peak, in terms of2-theta, selected from about 13.5, about 14.7, about 16.7, about 18.6,about 19.3, about 20.0, about 20.7, about 22.4, about 25.7, about 26.8,about 27.2, and about 28.1 degrees. In some embodiments, the crystallinesolid of Compound 1 methanesulfonic acid salt has at least twocharacteristic XRPD peaks, in terms of 2-theta, selected from about13.5, about 14.7, about 16.7, about 18.6, about 19.3, about 20.0, about20.7, about 22.4, about 25.7, about 26.8, about 27.2, and about 28.1degrees. In some embodiments, the crystalline solid of Compound 1methanesulfonic acid salt has at least three characteristic XRPD peaks,in terms of 2-theta, selected from about 13.5, about 14.7, about 16.7,about 18.6, about 19.3, about 20.0, about 20.7, about 22.4, about 25.7,about 26.8, about 27.2, and about 28.1 degrees.

The present invention further provides a (1 S)-(+)-10-camphorsulfonicacid salt of Compound 1. In some embodiments, a solid form of the(1S)-(+)-10-camphorsulfonic acid salt has an XRPD pattern as shown onFIG. 42. In some embodiments, the (1S)-(+)-10-camphorsulfonic acid saltof Compound 1 is a crystalline solid. In some embodiments, thecrystalline solid of Compound 1 (1S)-(+)-10-camphorsulfonic acid salthas at least one characteristic XRPD peak, in terms of 2-theta, selectedfrom about 7.1, about 10.9, about 13.6, about 16.1, about 17.7, about18.8, about 19.9, and about 23.2 degrees. In some embodiments, thecrystalline solid of Compound 1 (1S)-(+)-10-camphorsulfonic acid salthas at least two characteristic XRPD peaks, in terms of 2-theta,selected from about 7.1, about 10.9, about 13.6, about 16.1, about 17.7,about 18.8, about 19.9, and about 23.2 degrees. In some embodiments, thecrystalline solid of Compound 1 (1S)-(+)-10-camphorsulfonic acid salthas at least three characteristic XRPD peaks, in terms of 2-theta,selected from about 7.1, about 10.9, about 13.6, about 16.1, about 17.7,about 18.8, about 19.9, and about 23.2 degrees.

The present invention further provides a fumaric acid salt. In someembodiments, a solid form of the fumaric acid salt has an XRPD patternas shown on FIG. 43. In some embodiments, the fumaric acid salt isamorphous.

The present invention further provides a sulfuric acid salt ofCompound 1. In some embodiments, a solid form of the sulfuric acid salthas an XRPD pattern as shown on FIG. 44. In some embodiments, thesulfuric acid salt is amorphous.

The present invention further provides an L-tartaric acid salt ofCompound 1. In some embodiments, the L-tartaric acid salt is amorphous.A solid form of the L-tartaric acid salt has an XRPD pattern shown inFIG. 45.

The present invention further provides a D-tartaric acid salt ofCompound 1. In some embodiments, the D-tartaric acid salt is amorphous.A solid form of the L-tartaric acid salt has an XRPD pattern shown inFIG. 46.

Synthetic Preparation of Phosphoric Acid Salts

Generally, the phosphoric acid salts of the invention can be prepared bycombiningN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(referred to herein as “Compound 1” or “Compound 1 free base”) withphosphoric acid. In some embodiments, the phosphoric acid is provided inmolar excess relative to Compound 1 free base. In some embodiments, thecombining of Compound 1 free base and phosphoric acid is carried out inthe presence of a solvent. In some embodiments, the solvent compriseswater, methanol, 2-propanol, or a mixture thereof. In some embodiments,the combining can be carried out at elevated temperature such as, forexample, about 40 to about 80, about 50 to about 70, or about 55 toabout 65° C. In some embodiments, the Compound 1 phosphoric acid saltproduct obtained from the combining is substantially crystalline. Insome embodiments, the crystalline product comprises one or more of FormsI, II, III, IV, V, and VI. In some embodiments, the crystalline productcomprises Form I. In some embodiments, the crystalline productsubstantially comprises Form I. In some embodiments, the Compound 1phosphoric acid salt product obtained from the combining of phosphoricacid with Compound 1 is substantially amorphous or contains amorphoussolid.

Compound 1 free base, a precursor to the phosphate salt, can be preparedby combiningN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamidedihydrochloric acid salt (referred to herein as “Compound 1dihydrochloric acid salt” or “Compound 1 dihydrochloride”) with sodiumcarbonate. In some embodiments, the sodium carbonate is provided inmolar excess with respect to Compound 1 dihydrochloride. In someembodiments, the combining of Compound 1 dihydrochloric acid and sodiumcarbonate is carried out in the presence of solvent. In someembodiments, the solvent comprises water, methylene chloride, or amixture thereof. In some embodiments, the combining is carried out atroom temperature or at elevated temperature. Example reactiontemperatures include about 20 to about 40, about 20 to about 30, andabout 23 to about 27° C.

Compound 1 dihydrochloric acid salt can be prepared by reactingtert-butyl{(3R,4R,5S)-4-{[tert-butyl(dimethyl)silyl]oxy}-1-[(7R)-7-{[tert-butyl(dimethyl)silyl]oxy}-3-({[6-(2,6-difluorophenyl)-5-fluoropyridin-2-yl]carbonyl}amino)-6,7-dihydro-5H-cyclopenta[b]pyridin-4-yl]-5-methylpiperidin-3-yl}carbamate(25):

with hydrogen chloride. In some embodiments, the hydrogen chloride isprovided in molar excess such as, for example, between about 2 and about30, between about 5 and about 25, between about 10 and about 20, orabout 15 equivalents with respect to (25). In some embodiments, thereacting with hydrogen chloride is carried out in the presence of asolvent. In some embodiments, the solvent comprises 1,4-dioxane,methanol, or a mixture thereof. In some embodiments, the reaction withhydrogen chloride is carried out at room temperature.

The intermediate (25) can be prepared by coupling tert-butyl((3R,4R,5S)-1-(3-amino-7-{[tert-butyl(dimethyl)silyl]oxy}-6,7-dihydro-5H-cyclopenta[b]pyridin-4-yl)-4-{[tert-butyl(dimethyl)silyl]oxy}-5-methylpiperidin-3-yl)carbamate:

with 6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxylic acid:

In some embodiments, (24) is provided in slight molar excess (e.g.,about 1.1 to about 1.5 eq, or about 1.2 eq) with respect to (23). Insome embodiments, the coupling is carried out in the presence ofN,N-diisopropylethylamine (DIEA) andN,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uroniumhexafluorophosphate (HATU). In some embodiments, the DIEA is provided inmolar excess with respect to (23) (e.g., about 3 to 6 eq, or about 5eq). In some embodiments, the HATU is provided in molar excess withrespect to (23) (e.g., about 1.5 to 3.5 eq, or about 2.4 eq). In furtherembodiments, the coupling is carried out in the presence of a solvent.In some embodiments, the solvent comprises dimethylformamide (DMF). Insome embodiments, the coupling is carried out at about 10 to about 40,or about 15 to about 30° C.

The intermediate (23) can be prepared by mixing tert-butyl((3R,4R,5S)-1-((7R)-3-(aminocarbonyl)-7-{[tert-butyl(dimethyl)silyl]oxy}-6,7-dihydro-5H-cyclopenta[b]pyridin-4-yl)-4-{[tert-butyl(dimethyl)silyl]oxy}-5-methylpiperidin-3-yl)carbamate:

with tetra-N-butylammonium bromide (TBAB) in the presence of sodiumhydroxide. In some embodiments, the TBAB is provided in an amount ofabout 1 eq with respect to (22). In some embodiments, the sodiumhydroxide is provided in molar excess (e.g., about 1.1 to about 7 eq, orabout 2 to about 4 eq, or about 4.5 eq with respect to (22). In someembodiments, the mixing is carried out in the presence of1,3-dibromo-5,5-dimethylhydantoin. In some embodiments, the amount of1,3-dibromo-5,5-dimethylhydantoin provided is less than 1 eq (e.g.,about 0.1 to about 0.9 eq, or about 0.3 to about 0.8 eq, or about 0.7eq) with respect to (22). In some embodiments, the mixing is carried outin the presence of solvent. In some embodiments, the solvent comprisestetrahydrofuran (THF). In some embodiments, the mixing is carried out ata temperature which is below room temperature, such as at about 0 toabout 20, about 0 to about 15, or about 5 to about 10° C.

The intermediate (22) can be prepared by reacting tert-butyl[(3R,4R,5S)-4-{[tert-butyl(dimethyl)silyl]oxy}-1-((7R)-7-{[tert-butyl(dimethyl)silyl]oxy}-3-cyano-6,7-dihydro-5H-cyclopenta[b]pyridin-4-yl)-5-methylpiperidin-3-yl]carbamate:

with acetaldoxime. In some embodiments, the reacting with acetaldoximeis carried out in the presence of a palladium catalyst such as[1,1′-bis(diphenylphosphino)ferrocene]-dichloropalladium(II)(Pd(dppf)₂Cl₂complexed with dichloromethane. In some embodiments, the acetaldoxime isprovided in molar excess (e.g., about 2 to about 20 eq, about 5 to about15 eq, or about 10 eq). In some embodiments, the total amount ofacetaldoxime is delivered to the reaction mixture in portions. In someembodiments, the reacting with acetaldoxime is carried out in thepresence of a solvent. In some embodiments, the solvent comprises water,ethanol, or a mixture thereof. In some embodiments, the reacting iscarried out at elevated temperature (e.g., about 50 to about 150, about70 to about 100, or about 90° C.).

The intermediate (21) can be prepared by reacting(7R)-7-{[tert-butyl(dimethyl)silyl]oxy}-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carbonitrile:

with tert-butyl((3R,4R,5S)-4-{[tert-butyl(dimethyl)silyl]oxy}-5-methylpiperidin-3-yl)carbamate(9), or hydrochloric acid salt thereof (9 HCl):

in the presence of N,N-diisopropylethylamine (DIEA). In someembodiments, intermediate (9) is provided in slight molar excess (e.g.,1.05 eq) relative to (20). In some embodiments, the DIEA is provided inmolar excess (e.g., about 2 to about 6 eq, or about 4 eq) relative to(20). In some embodiments, the reacting of (20) with (9) is carried outin the presence of a solvent. In some embodiments, the solvent comprisesdimethylsulfoxide. In some embodiments, the reacting is carried out atroom temperature or at elevated temperature (e.g., at about 30 to about150, or at about 100° C.).

The intermediate(R)-7-(tert-butyldimethylsilyloxy)-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carbonitrile(20) can be prepared by reacting(R)-7-(tert-butyldimethylsilyloxy)-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carbaldehyde(19):

with ammonia and iodine. In some embodiments, the reacting is carriedout in the presence of a solvent. In some embodiments the solventcomprises water, THF, or a mixture thereof. In some embodiments thereacting is carried out at a temperature below room temperature, forexample between about 10 and 22° C. In some embodiments, the ammonia andiodine are provided in molar excess.

The intermediate(R)-7-(tert-butyldimethylsilyloxy)-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carbaldehyde(19) can be prepared by combining(R)-7-(tert-butyldimethylsilyloxy)-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine(18):

with n-butyl lithium in the presence of 2,2,6,6-tetramethylpiperidinefollowed by adding N,N-dimethylformamide (DMF). In some embodiments, thecombining is carried out below room temperature such as, for example,from −100 to −10° C. In some embodiments, the combining is carried outin a solvent. In some embodiments, the solvent comprises THF, hexane, ora mixture thereof.

The intermediate(R)-7-(tert-butyldimethylsilyloxy)-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine(18) can be prepared by reacting(R)-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-7-ol (17):

with tert-butyldimethylsilyl chloride and 1H-imidazole. In someembodiments, the reacting is carried out at a temperature below roomtemperature, such as at about −15 to 15 or about −15 to 0° C. In someembodiments, the reacting is carried out in the presence of a solventsuch as methylene chloride or other organic solvent. In someembodiments, the tert-butyldimethylsilyl chloride is provided in anamount of about 1 equivalent with respect to (17). In some embodiments,the 1H-imidazole is provided in molar excess with respect to (17).

The intermediate (R)-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-7-ol(17) can be prepared by reacting4-chloro-5H-cyclopenta[b]pyridin-7(6H)-one (16):

with formic acid in the presence of RuCl(p-cymene)[(R,R)-Ts-DPEN] andtriethylamine (TEA). In some embodiments, the reacting is carried out inthe presence of a solvent such as methylene chloride or other organicsolvent. In some embodiments, the reacting is carried out at atemperature below room temperature. In some embodiments, theRuCl(p-cymene)[(R,R)-Ts-DPEN] is provided in a catalytic amount. In someembodiments, the TEA is provided in molar excess relative to (16). Insome embodiments, the formic acid is provided in molar excess relativeto (16).

The intermediate 4-chloro-5H-cyclopenta[b]pyridin-7(6H)-one (16) can beprepared by reacting 4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-7-ol(15):

with pyridine-sulfur trioxide in the presence ofN,N-diisopropylethylamine. In some embodiments, (15) andN,N-diisopropylethylamine are combined prior to addition ofpyridine-sulfur trioxide. In some embodiments, the reacting is carriedout in the presence of solvent. In some embodiments, the solventcomprises methylene chloride or other organic solvent. In someembodiments, the reaction is carried out below room temperature, such asat about 0° C. In some embodiments, the N,N-diisopropylethylamine isprovided in molar excess relative to (15). In some embodiments, thepyridine-sulfur trioxide is provided in molar excess relative to (15).

The intermediate 4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-7-ol (15)can be prepared by reacting4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-7-yl acetate (14):

wherein Ac is acetyl, with potassium carbonate. In some embodiments, thereacting is carried out in the presence of solvent such as methanol,water, other polar solvent, or mixture thereof. In some embodiments, thereacting is carried out below room temperature, such as at about 0° C.In some embodiments, the potassium carbonate is provided in molar excessrelative to (14).

The intermediate 4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-7-ylacetate (14) can be prepared by reacting4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine 1-oxide (13):

with acetic anhydride. In some embodiments, the reacting is carried outin the presence of an organic solvent, wherein the organic solventcomprises toluene or other non-polar solvent. In some embodiments, thereacting is carried out at elevated temperature, such as at about 50 toabout 150, or about 70 to about 90, or about 80 to about 85° C. In someembodiments, the acetic anhydride is provided in molar excess relativeto (13).

The intermediate 4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine 1-oxide(13) can be prepared by reacting4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine (12):

with urea hydrogen peroxide (UHP) in the presence of a catalyst. In someembodiments, the catalyst is a transition metal catalyst such asmethyltrioxorhemium(VII). In some embodiments, the catalyst is providedin a catalytic amount (e.g., <0.1 eq relative to (12)). In someembodiments, the reacting is carried out in the presence of solvent. Insome embodiments, the solvent comprises methanol or other polar solvent.In some embodiments, the reacting is carried out at room temperature. Insome embodiments, the UHP is provided in an amount of about 1 to about 2equivalents, or in an amount of about 1.5 equivalents with respect to(12).

The intermediate 4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine (12) canbe prepared by reacting 6,7-dihydro-5H-cyclopenta[b]pyridine 1-oxide(11):

with phosphoryl chloride. In some embodiments, the phosphoryl chlorideis provided in an amount of about 2-4 equivalents, or about 3equivalents with respect to (11). In some embodiments, the reacting iscarried out in the presence of an organic solvent, where the organicsolvent comprise, for example, toluene. In some embodiments, thereacting is carried out at elevated temperature such as at about 50 toabout 100, or about 70 to about 90, or about 80 to about 85° C.

The intermediate 6,7-dihydro-5H-cyclopenta[b]pyridine 1-oxide (11) canbe prepared by reacting 6,7-dihydro-5H-cyclopenta[b]pyridine (10):

with urea hydrogen peroxide UHP in the presence of a catalyst. In someembodiments, the catalyst is a transition metal catalyst such asmethyltrioxorhenium(VII). In some embodiments, the catalyst is providedin a catalytic amount (e.g., <0.1 eq relative to (10)). In someembodiments, the reacting is carried out at about room temperature. Insome embodiments the UHP is provided in an amount of about 2-4 eq orabout 3 eq with respect to (10). In some embodiments, the reacting iscarried out in the presence of solvent, such as a solvent comprisingmethanol or other polar organic solvent. In some embodiments, thepresent invention relates to an intermediate compound selected from:

wherein TBS is tert-butyl(dimethyl)silyl.

In some embodiments, the present invention relates to an intermediatecompound selected from:

wherein TBS is tert-butyl(dimethyl)silyl and Boc istert-butyloxycarbonyl.

In some embodiments, the intermediate compound is

In some embodiments, the intermediate compound is

In some embodiments, the intermediate compound is

wherein TBS is tert-butyl(dimethyl)silyl.

In some embodiments, the intermediate compound is

wherein TBS is tert-butyl(dimethyl)silyl.

In some embodiments, the intermediate compound is

wherein TBS is tert-butyl(dimethyl)silyl.

In some embodiments, the intermediate compound is

wherein TBS is tert-butyl(dimethyl)silyl and Boc istert-butyloxycarbonyl.

In some embodiments, the intermediate compound is

wherein TBS is tert-butyl(dimethyl)silyl and Boc istert-butyloxycarbonyl.

In some embodiments, the intermediate compound is

wherein TBS is tert-butyl(dimethyl)silyl and Boc istert-butyloxycarbonyl.

In some embodiments, the intermediate compound is

wherein TBS is tert-butyl(dimethyl)silyl and Boc istert-butyloxycarbonyl.

In some embodiments, the present invention provides a method ofpreparing Compound 1 phosphoric acid salt, comprising:

-   -   reacting Compound 10 with urea hydrogen peroxide and        methyltrioxorhenium(VII) to form Compound 11;    -   reacting Compound 11 with phosphoryl chloride to form Compound        12;    -   reacting Compound 12 with urea hydrogen peroxide and        methyltrioxorhemium(VII) to form Compound 13;    -   reacting Compound 13 with acetic anhydride to form Compound 14;    -   reacting Compound 14 with potassium carbonate to form Compound        15;    -   reacting Compound 15 with pyridine-sulfur trioxide in the        presence of N,N-diisopropylethylamine to form Compound 16;    -   reacting Compound 16 with formic acid in the presence of        RuCl(p-cymene)[(R,R)-Ts-DPEN] and triethylamine (TEA) to form        Compound 17;    -   reacting Compound 17 with tert-butyldimethylsilyl chloride and        1H-imidazole to form Compound 18;    -   combining Compound 18 with n-butyl lithium in the presence of        2,2,6,6-tetramethylpiperidine followed by adding        N,N-dimethylformamide (DMF) to form Compound 19;    -   reacting Compound 19 with ammonia and iodine to form Compound        20;    -   reacting Compound 20 with Compound 9, or hydrochloric acid salt        thereof, in the presence of N,N-diisopropylethylamine (DIEA) to        form Compound 21;    -   reacting compound 21 with acetaldoxime to form Compound 22;    -   mixing Compound 22 with tetra-N-butylammonium bromide (TBAB) in        the presence of sodium hydroxide to form Compound 23;    -   coupling Compound 23 with Compound 24 to form Compound 25;

reacting Compound 25 with hydrogen chloride to form Compound 1dihydrochloric acid salt;

combining Compound 1 dihydrochloric acid salt with sodium carbonate toform Compound 1 free base; and

combining Compound 1 free base with phosphoric acid to form Compound 1phosphoric acid salt.

In some embodiments, the present invention provides a method ofpreparing Compound 1 phosphoric acid salt, comprising:

reacting Compound 19 with ammonia and iodine to form Compound 20;

reacting Compound 20 with Compound 9, or hydrochloric acid salt thereof,in the presence of N,N-diisopropylethylamine (DIEA) to form Compound 21;

reacting compound 21 with acetaldoxime to form Compound 22;

mixing Compound 22 with tetra-N-butylammonium bromide (TBAB) in thepresence of sodium hydroxide to form Compound 23;

coupling Compound 23 with Compound 24 to form Compound 25;

reacting Compound 25 with hydrogen chloride to form Compound 1dihydrochloric acid salt;

combining Compound 1 dihydrochloric acid salt with sodium carbonate toform Compound 1 free base; and

combining Compound 1 free base with phosphoric acid to form Compound 1phosphoric acid salt.

In some embodiments, the present invention provides a method ofpreparing Compound 20, comprising:

reacting Compound 10 with urea hydrogen peroxide andmethyltrioxorhenium(VII) to form Compound 11;

reacting Compound 11 with phosphoryl chloride to form Compound 12;

reacting Compound 12 with urea hydrogen peroxide andmethyltrioxorhemium(VII) to form Compound 13;

reacting Compound 13 with acetic anhydride to form Compound 14;

reacting Compound 14 with potassium carbonate to form Compound 15;

reacting Compound 15 with pyridine-sulfur trioxide in the presence ofN,N-diisopropylethylamine to form Compound 16;

reacting Compound 16 with formic acid in the presence ofRuCl(p-cymene)[(R,R)-Ts-DPEN] and triethylamine (TEA) to form Compound17;

reacting Compound 17 with tert-butyldimethylsilyl chloride and1H-imidazole to form Compound 18;

combining Compound 18 with n-butyl lithium in the presence of2,2,6,6-tetramethylpiperidine followed by adding N,N-dimethylformamide(DMF) to form Compound 19; and

reacting Compound 19 with ammonia and iodine to form Compound 20.

As used herein, the term “reacting” is used as known in the art andgenerally refers to the bringing together of chemical reagents in such amanner so as to allow their interaction at the molecular level toachieve a chemical or physical transformation. In some embodiments, thereacting involves at least two reagents. In some embodiments, thereacting step of a synthetic process may involve one or more substancesin addition to the reagents such as solvent and/or a catalyst. Thereacting steps of the processes described herein can be conducted for atime and under conditions suitable for preparing the identified product.The terms “combining” and “mixing” with respect to reagents of achemical reaction are used interchangeably with the term “reacting”herein. The term “coupling” also can be considered interchangeable with“reacting” but may be used in conjunction with a reaction step thatinvolves the linking of two organic fragments.

The processes described herein can be monitored according to anysuitable method known in the art. For example, product formation can bemonitored by spectroscopic means, such as nuclear magnetic resonancespectroscopy (e.g., ¹H or ¹³C), infrared spectroscopy, spectrophotometry(e.g., UV-visible), or mass spectrometry; or by chromatography such ashigh performance liquid chromatography (HPLC) or thin layerchromatography. The compounds obtained by the reactions can be purifiedby any suitable method known in the art. For example, chromatography(medium pressure) on a suitable adsorbent (e.g., silica gel, alumina andthe like), HPLC, or preparative thin layer chromatography; distillation;sublimation, trituration, or recrystallization. The purity of thecompounds, in general, are determined by physical methods such asmeasuring the melting point (in case of a solid), obtaining a NMRspectrum, or performing a HPLC separation. If the melting pointdecreases, if unwanted signals in the NMR spectrum are decreased, or ifextraneous peaks in an HPLC trace are removed, the compound can be saidto have been purified. In some embodiments, the compounds aresubstantially purified.

Preparation of compounds can involve the protection and deprotection ofvarious chemical groups. The need for protection and deprotection, andthe selection of appropriate protecting groups can be readily determinedby one skilled in the art. The chemistry of protecting groups can befound, for example, in Wuts and Greene, Greene's Protective Groups inOrganic Synthesis, 4^(th) Ed., John Wiley & Sons: New York, 2006, whichis incorporated herein by reference in its entirety.

The reactions of the processes described herein can be carried out atappropriate temperatures which can be readily determined by the skilledartisan. Reaction temperatures will depend on, for example, the meltingand boiling points of the reagents and solvent, if present; thethermodynamics of the reaction (e.g., vigorously exothermic reactionsmay need to be carried out at reduced temperatures); and the kinetics ofthe reaction (e.g., a high activation energy barrier may need elevatedtemperatures). “Elevated temperature” refers to temperatures above roomtemperature (about 22° C.).

The reactions of the processes described herein can be carried out insuitable solvents which can be readily selected by one of skill in theart of organic synthesis. Suitable solvents can be substantiallynon-reactive with the starting materials (reactants), the intermediates,or products at the temperatures at which the reactions are carried out,i.e., temperatures which can range from the solvent's freezingtemperature to the solvent's boiling temperature. A given reaction canbe carried out in one solvent or a mixture of more than one solvent.Depending on the reaction step, suitable solvent(s) for that particularreaction step can be selected. Appropriate solvents include water,alkanes (such as pentanes, hexanes, heptanes, cyclohexane, etc., or amixture thereof), aromatic solvents (such as benzene, toluene, xylene,etc.), alcohols (such as methanol, ethanol, isopropanol, etc.), ethers(such as dialkylethers, methyl tert-butyl ether (MTBE), tetrahydrofuran(THF), dioxane, etc.), esters (such as ethyl acetate, butyl acetate,etc.), halogenated hydrocarbon solvents (such as dichloromethane (DCM),chloroform, dichloroethane, tetrachloroethane), dimethylformamide (DMF),dimethylsulfoxide (DMSO), acetone, acetonitrile (ACN),hexamethylphosphoramide (HMPA) and N-methyl pyrrolidone (NMP). Suchsolvents can be used in either their wet or anhydrous forms.

Resolution of racemic mixtures of compounds can be carried out by any ofnumerous methods known in the art. An example method includes fractionalrecrystallization using a “chiral resolving acid” which is an opticallyactive, salt-forming organic acid. Suitable resolving agents forfractional recrystallization methods are, for example, optically activeacids, such as the D and L forms of tartaric acid, diacetyltartaricacid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid orthe various optically active camphorsulfonic acids. Resolution ofracemic mixtures can also be carried out by elution on a column packedwith an optically active resolving agent (e.g.,dinitrobenzoylphenylglycine). Suitable elution solvent composition canbe determined by one skilled in the art.

Preparation of Phosphate Salt Crystalline Forms I-VI

Form I can be prepared, for example, by precipitating the solid formfrom a solution comprisingN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamidephosphoric acid salt. In some embodiments, the solution comprises asolvent that comprises 2-propanol. In other embodiments, the solutioncomprises a solvent that comprises acetonitrile and/or ethanol. Theprecipitating can be carried out at any suitable temperature, such asabout room temperature or at elevated temperature. The precipitating canbe carried out by any means which concentrates the solution, such as byevaporation, addition of anti-solvent, or cooling.

Form II can be prepared, for example, by precipitating the solid formfrom a solution ofN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamidephosphoric acid salt in a solvent comprising dimethylformamide (DMF).The precipitating can be carried out by any means which concentrates thesolution, such as by evaporation, addition of anti-solvent, or cooling.In some embodiments, the precipitating is carried out by evaporationunder air at about room temperature (e.g., about 25° C.).

Form III can be prepared by precipitating the solid form from a solutionofN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamidephosphoric acid salt in a solvent comprising dimethylformamide (DMF),wherein the precipitating can be carried out at elevated temperature,such as at about 30° C. to about 70° C., at about 40° C. to about 60°C., or at about 50° C.

Form IV can be prepared by precipitating the solid form from an aqueoussolution ofN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamidephosphoric acid salt. In some embodiments, the precipitating is carriedout by evaporation under air at elevated temperature such as, forexample, between about 30° C. and about 70° C., between about 40° C. andabout 60° C., or between about 45° C. and about 55° C.

Form V can be prepared by precipitating the solid form from an aqueoussolution ofN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamidephosphoric acid salt by cooling the solution. In some embodiments, theaqueous solution is at a temperature of about 30 to about 40° C., orabout 35° C. prior to cooling. In some embodiments, the solution iscooled to about 4-5° C. In some embodiments, the cooling is carried outby quench-cooling.

Form VI can be prepared by filtering a slurry ofN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamidephosphoric acid salt in water to obtain a filtrate and then cooling thefiltrate. (e.g., to below about 10° C., or to about 4-5° C.).

Synthetic Preparation of Other Salt Forms

The dihydrochloric acid salt can be prepared, for example, by combiningN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1 free base) with hydrochloric acid. In some embodiments, thecombining is carried out in the presence of a solvent, such as a solventcomprising 2-propanol and/or 2-propyl acetate. In some embodiments, thehydrochloric acid is provided in molar excess with respect to Compound 1free base. In some embodiments, the molar ratio of Compound 1 free baseto hydrochloric acid is from about 1:2 to about 1:2.5. In someembodiments, the ratio of Compound 1 free base to hydrochloric acid isabout 1:2.34.

The monohydrochloric acid salt can be prepared, for example, bycombiningN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1 free base) with hydrochloric acid. In some embodiments, thecombining is carried out in the presence of solvent such as a solventcomprising 2-propanol and/or 2-propyl acetate. In some embodiments, themolar ratio of Compound 1 free base to hydrochloric acid is from about1:1 to about 1:1.5. In some embodiments, the molar ratio of Compound 1free base to hydrochloric acid is about 1:1.12.

The maleic acid salt can be prepared, for example, by combiningN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1 free base) with maleic acid. In some embodiments, combiningis carried out in the presence of a solvent such as a solvent comprising2-propanol. In some embodiments, the molar ratio of Compound 1 free baseto maleic acid is from about 1:1 to about 1:1.5. In some embodiments,the molar ratio of Compound 1 free base to maleic acid is about 1:1.21.In some embodiments, the method comprises adding seed crystals to induceprecipitation.

The adipic acid salt can be prepared, for example, by combiningN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1 free base) with adipic acid. In some embodiments, thecombining is carried out in the presence of a solvent such as a solventcomprising 2-propanol and/or heptane. In some embodiments, the molarratio of Compound 1 free base to adipic acid is from about 1:2 to about1:3. In some embodiments, the molar ratio of Compound 1 free base toadipic acid is about 1:2.49.

Hydrobromic acid salts can be prepared, for example, by combiningN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1 free base) with hydrobromic acid. In some embodiments, thecombining is carried out in the presence of a solvent such as a solventcomprising 2-propanol and/or water. In some embodiments, the molar ratioof Compound 1 free base to hydrobromic acid is from about 1:2 to about1:3 during the combining. In some embodiments, the molar ratio ofCompound 1 free base to hydrobromic acid is about 1:2.4. In someembodiments, the hydrobromic acid salt is a dihydrobromic acid salt. Infurther embodiments, the hydrobromic acid salt is a monohydrobromic acidsalt.

The mandelic acid salt can be prepared, for example, by combiningN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1 free base) with (R)-(−)-mandelic acid. In some embodiments,the combining is carried out in the presence solvent, such as a solventcomprising 2-propanol. In some embodiments, the molar ratio of Compound1 free base to (R)-(−)-mandelic acid is from about 1:1 to about 1:1.5during the combining. In some embodiments, the molar ratio of Compound 1free base to (R)-(−)-mandelic acid is about 1:1.06.

The salicylic acid salt can be prepared, for example, by combiningN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1 free base) with salicylic acid, optionally in the presenceof a solvent. In some embodiments, the solvent comprises isopropylalcohol. In some embodiments, the acid is provided in molar excess withrespect to Compound 1 free base. In some embodiments, the molar ratio ofCompound 1 free base to salicylic acid is from about 1:1 to about 1:1.5or about 1:1 to about 1:1.2. In some embodiments, the molar ratio ofCompound 1 free base to salicylic acid is about 1:1.16. In someembodiments, the combining is carried out at about room temperature.

The benzoic acid salt can be prepared, for example, by combiningN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1 free base) with benzoic acid, optionally in the presence ofa solvent. In some embodiments, the solvent comprises isopropyl alcohol.In some embodiments, the acid is provided in molar excess with respectto Compound 1 free base. In some embodiments, the molar ratio ofCompound 1 free base to benzoic acid is from about 1:1 to about 1:1.5 orabout 1:1 to about 1:1.2. In some embodiments, the molar ratio ofCompound 1 free base to benzoic acid is about 1:1.16. In someembodiments, the combining is carried out at about room temperature.

The benzenesulfonic acid salt can be prepared, for example, by combiningN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1 free base) with benzenesulfonic acid optionally in thepresence of a solvent. In some embodiments, the solvent comprisesisopropyl alcohol. In some embodiments, the acid is provided in molarexcess with respect to Compound 1 free base. In some embodiments, themolar ratio of Compound 1 free base to benzenesulfonic acid is fromabout 1:1 to about 1:1.5 or about 1:1 to about 1:1.2. In someembodiments, the molar ratio of Compound 1 free base to benzenesulfonicacid is about 1:1.1. In some embodiments, the combining is carried outat about room temperature.

The L-pyroglutamic acid salt can be prepared, for example, by combiningN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1 free base) with L-pyroglutamic acid optionally in thepresence of a solvent In some embodiments, the solvent comprisesisopropyl alcohol. In some embodiments, the solvent comprises isopropylalcohol and heptane. In some embodiments, the acid is provided in molarexcess with respect to Compound 1 free base. In some embodiments, themolar ratio of Compound 1 free base to L-pyroglutamic acid is from about1:1 to about 1:1.5 or about 1:1 to about 1:1.2. In some embodiments, themolar ratio of Compound 1 free base to L-pyroglutamic acid is about1:1.12. In some embodiments, the combining is carried out at about roomtemperature.

The methanesulfonic acid salt can be prepared, for example, by combiningN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1 free base) with methanesulfonic acid optionally in thepresence of a solvent. In some embodiments, the solvent comprisesisopropyl alcohol. In some embodiments, the solvent comprises isopropylalcohol and ethanol. In some embodiments, the solvent comprisesisopropyl alcohol, ethanol, and heptane. In some embodiments, the acidis provided in molar excess with respect to Compound 1 free base. Insome embodiments, the molar ratio of Compound 1 free base tomethanesulfonic acid is from about 1:1 to about 1:1.5 or about 1:1 toabout 1:1.2. In some embodiments, the molar ratio of Compound 1 freebase to methanesulfonic acid is about 1:1.1. In some embodiments, thecombining is carried out at about room temperature.

The (1S)-(+)-10-camphorsulfonic acid salt can be prepared, for example,by combiningN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1 free base) with (1S)-(+)-10-camphorsulfonic acid optionallyin the presence of a solvent. In some embodiments, the solvent comprisesisopropyl alcohol. In some embodiments, the solvent comprises isopropylalcohol and heptane. In some embodiments, the acid is provided in molarexcess with respect to Compound 1 free base. In some embodiments, themolar ratio of Compound 1 free base to (1S)-(+)-10-camphorsulfonic acidis from about 1:1 to about 1:1.5 or about 1:1 to about 1:1.2. In someembodiments, the molar ratio of Compound 1 free base to(1S)-(+)-10-camphorsulfonic acid is about 1:1.1. In some embodiments,the combining is carried out at about room temperature.

The fumaric acid salt can be prepared, for example, by combiningN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1 free base) with fumaric acid optionally in the presence of asolvent. In some embodiments, the solvent comprises isopropyl alcohol.In some embodiments, the solvent comprises isopropyl alcohol andheptane. In some embodiments, the acid is provided in molar excess withrespect to Compound 1 free base. In some embodiments, the molar ratio ofCompound 1 free base to fumaric acid is about 1:1.16. In someembodiments, the combining is carried out at about room temperature.

The sulfuric acid salt can be prepared, for example, by combiningN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1 free base) with sulfuric acid optionally in the presence ofa solvent. In some embodiments, the solvent comprises isopropyl alcohol.In some embodiments, the acid is provided in molar excess with respectto Compound 1 free base. In some embodiments, the molar ratio ofCompound 1 free base to sulfuric acid is from about 1:1 to about 1:1.5or about 1:1 to about 1:1.2. In some embodiments, the molar ratio ofCompound 1 free base to sulfuric acid is about 1:1.1. In someembodiments, the combining is carried out at about room temperature.

The L-tartaric acid salt can be prepared, for example, by combiningN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1 free base) with L-tartaric acid optionally in the presenceof a solvent. In some embodiments, the solvent comprises isopropylalcohol. In some embodiments, the solvent comprises isopropyl alcoholand heptane. In some embodiments, the acid is provided in molar excesswith respect to Compound 1 free base. In some embodiments, the molarratio of Compound 1 free base to L-tartaric acid is from about 1:1 toabout 1:1.5 or about 1:1 to about 1:1.2. In some embodiments, the molarratio of Compound 1 free base to L-tartaric acid is about 1:1.16. Insome embodiments, the combining is carried out at about roomtemperature.

The D-tartaric acid salt can be prepared, for example, by combiningN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1 free base) with D-tartaric acid optionally in the presenceof a solvent. In some embodiments, the solvent comprises isopropylalcohol. In some embodiments, the solvent comprises isopropyl alcoholand heptane. In some embodiments, the acid is provided in molar excesswith respect to Compound 1 free base. In some embodiments, the molarratio of Compound 1 free base to D-tartaric acid is from about 1:1 toabout 1:1.5 or about 1:1 to about 1:1.2. In some embodiments, the molarratio of Compound 1 free base to D-tartaric acid is about 1:1.16. Insome embodiments, the combining is carried out at about roomtemperature.

Methods of Use

Compound 1 and the salts described herein can inhibit the activity ofone or more members of the Pim kinase family and, thus, is useful intreating diseases and disorders associated with activity of Pim kinases.For example, Compound 1 and its salts can inhibit one or more of Pim1,Pim2 and Pim3. Thus, the present disclosure provides methods of treatinga Pim kinase-associated disease or disorder in an individual (e.g.,patient) by administering to the individual in need of such treatment atherapeutically effective amount or dose of Compound 1 phosphoric acidsalt, or any of the embodiments thereof, or a pharmaceutical compositionthereof. The present disclosure also provides a Compound 1 phosphoricacid salt, or any of the embodiments thereof, or a pharmaceuticalcomposition thereof, for use in treating a Pim kinase-associated diseaseor disorder. Also provided is the use of Compound 1 phosphoric acidsalt, or any of the embodiments thereof, or a pharmaceutical compositionthereof, in the manufacture of a medicament for treating a Pimkinase-associated disease or disorder.

A Pim kinase-associated disease can include any disease, disorder orcondition that is directly or indirectly linked to expression oractivity of the Pim kinase, including over-expression and/or abnormalactivity levels. Abnormal activity levels can be determined by comparingactivity level in normal, healthy tissue or cells with activity level indiseased cells. A Pim kinase-associated disease can also include anydisease, disorder or condition that can be prevented, ameliorated,inhibited or cured by modulating Pim kinase activity. In someembodiments, the disease is characterized by the abnormal activity orexpression (e.g., overexpression) of one or more Pim1, Pim2 and Pim3. Insome embodiments, the disease is characterized by mutant Pim1, Pim2 orPim3. A Pim kinase associated disease can also refer to any disease,disorder or condition wherein modulating the expression or activity ofone or more Pim kinases is beneficial.

Pim kinase associated diseases that can be treated according to theinvention include cancer, including, in particular, cancers in which Pimkinases are upregulated or an oncogene, e.g., Myc or BCL2, is activated.Pim kinase associated diseases include solid tumors, e.g., prostatecancer, colon cancer, esophageal cancer, endometrial cancer, ovariancancer, uterine cancer, renal cancer, hepatic cancer, pancreatic cancer,gastric cancer, breast cancer, lung cancer, cancers of the head or neck,thyroid cancer, glioblastoma, sarcoma, bladder cancer, etc. Pim kinaseassociated diseases also include hematological cancers, e.g., lymphoma,leukemia such as acute lymphoblastic leukemia (ALL), acute myelogenousleukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenousleukemia (CML), diffuse large B-cell lymphoma (DLBCL), mantle celllymphoma, non-Hodgkin lymphoma (including relapsed non-Hodgkin lymphoma,refractory non-Hodgkin lymphoma and recurrent follicular non-Hodgkinlymphoma), Hodgkin lymphoma and multiple myeloma.

Pim kinase associated diseases that can be treated according to theinvention also include myeloproliferative disorders such as polycythemiavera (PV), essential thrombocythemia (ET), chronic myelogenous leukemia(CML) and the like. The myeloproliferative disorder can be myelofibrosissuch as primary myelofibrosis (PMF), post-polycythemia vera/essentialthrombocythemia myelofibrosis (Post-PV/ET MF), post-essentialthrombocythemia myelofibrosis (Post-ET MF) or post-polycythemia veramyelofibrosis (Post-PV MF).

Pim kinase-associated diseases that can be treated according to theinvention also include immune disorders such as autoimmune diseases. Theimmune disorders include multiple sclerosis, rheumatoid arthritis,allergy, food allergy, asthma, lupus, inflammatory bowel disease andulcerative colitis.

Pim kinase-associated diseases that can be treated according to theinvention also include atherosclerosis.

The salts of the invention can also be used to inhibit disease processesin which Pim-kinases are involved, including angiogenesis and tumormetastasis.

Due to the fact that Pim kinases are regulated by the JAK/STAT pathway,the salts of the invention are useful to treat diseases in whichmodulating JAK/STAT signaling is beneficial. Thus, other diseases thatcan be treated include Crohn's disease, irritable bowel syndrome,pancreatitis, diverticulosis, Grave's disease, juvenile rheumatoidarthritis, osteoarthritis, psoriatic arthritis, ankylosing spondylitis,myasthenia gravis, vasculitis, autoimmune thyroiditis, dermatitis,psoriasis, scleroderma, systemic sclerosis, vitiligo, graft versus hostdisease, Sjogren's syndrome, glomerulonephritis and diabetes mellitis(type I).

The terms “individual” or “patient,” used interchangeably, refer to anyanimal, including mammals, preferably mice, rats, other rodents,rabbits, dogs, cats, swine, cattle, sheep, horses, or primates, and mostpreferably humans.

The phrase “therapeutically effective amount” refers to the amount ofactive compound or pharmaceutical agent that elicits the biological ormedicinal response in a tissue, system, animal, individual or human thatis being sought by a researcher, veterinarian, medical doctor or otherclinician.

The term “treating” or “treatment” refers to one or more of (1)inhibiting the disease; e.g., inhibiting a disease, condition ordisorder in an individual who is experiencing or displaying thepathology or symptomatology of the disease, condition or disorder (i.e.,arresting further development of the pathology and/or symptomatology);and (2) ameliorating the disease; e.g., ameliorating a disease,condition or disorder in an individual who is experiencing or displayingthe pathology or symptomatology of the disease, condition or disorder(i.e., reversing the pathology and/or symptomatology) such as decreasingthe severity of disease.

In some embodiments, the salts of the present invention may be useful inpreventing or reducing the risk of developing the disease; e.g.,preventing or reducing the risk of developing a disease, condition ordisorder in an individual who may be predisposed to the disease,condition or disorder but does not yet experience or display thepathology or symptomatology of the disease.

Combination Therapies

Cancer cell growth and survival can be impacted by multiple signalingpathways. Thus, it is useful to combine different kinase inhibitors,exhibiting different preferences in the kinases which they modulate theactivities of, to treat such conditions. Targeting more than onesignaling pathway (or more than one biological molecule involved in agiven signaling pathway) may reduce the likelihood of drug-resistancearising in a cell population, and/or reduce the toxicity of treatment.

Accordingly, the Pim inhibitor of the present invention can be used incombination with one or more other kinase inhibitors for the treatmentof diseases, such as cancer, that are impacted by multiple signalingpathways. For example, a combination can include one or more inhibitorsof the following kinases for the treatment of cancer: Akt1, Akt2, Akt3,TGF-βR, PKA, PKG, PKC, CaM-kinase, phosphorylase kinase, MEKK, ERK,MAPK, mTOR, EGFR, HER2, HER3, HER4, INS-R, IGF-1R, IR-R, PDGFαR, PDGFβR,CSFIR, KIT, FLK-II, KDR/FLK-1, FLK-4, fit-1, FGFR1, FGFR2, FGFR3, FGFR4,c-Met, Ron, Sea, TRKA, TRKB, TRKC, FLT3, VEGFR/Flt2, Flt4, EphA1, EphA2,EphA3, EphB2, EphB4, Tie2, Src, Fyn, Lck, Fgr, Btk, Fak, SYK, FRK, JAK,ABL, ALK and B-Raf. Non-limiting examples of inhibitors that can becombined with the Pim inhibitor provided herein for treatment ofdiseases such as cancer include an FGFR inhibitor (FGFR1, FGFR2, FGFR3or FGFR4), a JAK inhibitor (JAK1 and/or JAK2, e.g., ruxolitinib,baricitinib or INCB39110), an IDO inhibitor (e.g., epacadostat andNLG919), a TDO inhibitor, a PI3K-delta inhibitor, a PI3K-gammainhibitor, a CSF1R inhibitor, a TAM receptor tyrosine kinases (Tyro-3,Axl, and Mer), an angiogenesis inhibitor, an interleukin receptorinhibitor and an adenosine receptor antagonist or combinations thereof.Additionally, the Pim inhibitors of the invention can be combined withinhibitors of kinases associated with the PIK3/Akt/mTOR signalingpathway, such as PI3K, Akt (including Akt1, Akt2 and Akt3) and mTORkinases.

In some embodiments Pim inhibitors of the invention can be combined withinhibitors selective for JAK1 and/or JAK2 (e.g., ruxolitinib,baricitinib, momelotinib, filgotinib, pacritinib, INCB039110,INCB052793, INCB054707, CYT387, ABT494, AZD1480, XL019, CEP-33779, AZ960, TG101209, and gandotinib). In some embodiments Pim inhibitors ofthe invention can be combined with inhibitors selective for JAK1 (e.g.INCB039110, INCB052793, INCB054707, and ABT494) such as those disclosedin e.g., WO 2010/135650, WO 2011/028685, WO 2011/112662, WO 2012/068450,WO 2012/068440, WO 2012/177606, WO 2013/036611, WO 2013/026025, WO2014/138168, WO 2013/173720, WO 2015/021153, WO 2014/071031, WO2014/106706, WO 2015/131031, WO 2015/168246, and WO 2015/184305. In someembodiments Pim inhibitors of the invention can be combined withinhibitors selective for JAK2 (e.g., pacritinib, AZD1480, XL019,CEP-33779, AZ 960, TG101209, and gandotinib).

In some embodiments Pim inhibitors of the invention can be combined withinhibitors selective for PI3K delta (e.g., idelalisib, INCB040093,INCB050465, and TGR 1202) such as those disclosed in e.g., WO2011/0008487, WO 2011/075643, WO 2011/075630, WO 2011/163195, WO2011/130342, WO 2012/087881, WO 2012/125629, WO 2012/135009, WO2013/033569, WO2013/151930, WO 2014/134426, WO 2015/191677, and WO2015/157257.

The Pim inhibitor of the present invention can further be used incombination with other methods of treating cancers, for example bychemotherapy, irradiation therapy, tumor-targeted therapy, adjuvanttherapy, immunotherapy, or surgery. Examples of immunotherapy includecytokine treatment (e.g., interferons, GM-CSF, G-CSF, IL-2), CRS-207immunotherapy, cancer vaccine, monoclonal antibody, adoptive T celltransfer, oncolytic virotherapy and immunomodulating small molecules,including thalidomide or JAK1/2 inhibitor and the like. For example, thesalts of the invention can be administered in combination with one ormore anti-cancer drugs, such as a chemotherapeutics. Examplechemotherapeutics include any of: abarelix, aldesleukin, alemtuzumab,alitretinoin, allopurinol, altretamine, anastrozole, arsenic trioxide,asparaginase, azacitidine, bevacizumab, bexarotene, bleomycin,bortezombi, bortezomib, busulfan intravenous, busulfan oral,calusterone, capecitabine, carboplatin, carmustine, cetuximab,chlorambucil, cisplatin, cladribine, clofarabine, cyclophosphamide,cytarabine, dacarbazine, dactinomycin, dalteparin sodium, dasatinib,daunorubicin, decitabine, denileukin, denileukin diftitox, dexrazoxane,docetaxel, doxorubicin, dromostanolone propionate, eculizumab,epirubicin, erlotinib, estramustine, etoposide phosphate, etoposide,exemestane, fentanyl citrate, filgrastim, floxuridine, fludarabine,fluorouracil, fulvestrant, gefitinib, gemcitabine, gemtuzumabozogamicin, goserelin acetate, histrelin acetate, ibritumomab tiuxetan,idarubicin, ifosfamide, imatinib mesylate, interferon alfa 2a,irinotecan, lapatinib ditosylate, lenalidomide, letrozole, leucovorin,leuprolide acetate, levamisole, lomustine, meclorethamine, megestrolacetate, melphalan, mercaptopurine, methotrexate, methoxsalen, mitomycinC, mitotane, mitoxantrone, nandrolone phenpropionate, nelarabine,nofetumomab, oxaliplatin, paclitaxel, pamidronate, panitumumab,pegaspargase, pegfilgrastim, pemetrexed disodium, pentostatin,pipobroman, plicamycin, procarbazine, quinacrine, rasburicase,rituximab, ruxolitinib, sorafenib, streptozocin, sunitinib, sunitinibmaleate, tamoxifen, temozolomide, teniposide, testolactone, thalidomide,thioguanine, thiotepa, topotecan, toremifene, tositumomab, trastuzumab,tretinoin, uracil mustard, valrubicin, vinblastine, vincristine,vinorelbine, vorinostat and zoledronate.

In some embodiments Pim inhibitors of the invention can be combined withcytarabine.

Other anti-cancer agent(s) include antibody therapeutics such astrastuzumab (Herceptin), antibodies to costimulatory molecules such asCTLA-4, 4-1BB, PD-1 and PD-L1, or antibodies to cytokines (IL-10, TGF-β,etc.). The Pim inhibitors presented herein can further be used incombination with one or more checkpoint inhibitors (e.g., inhibitors ofan immune checkpoint molecule). Exemplary immune checkpoint inhibitorsinclude inhibitors against immune checkpoint molecules such as CD27,CD28, CD40, CD122, CD96, CD73, CD47, OX40, GITR, CSF1R, JAK (e.g., JAK1and/or JAK2), PI3K delta, PI3K gamma, TAM, arginase, CD137 (also knownas 4-1BB), ICOS, A2AR, B7-H3, B7-H4, BTLA, CTLA-4, LAG3, TIM3, VISTA,PD-1, PD-L1 and PD-L2. In some embodiments, the immune checkpointmolecule is a stimulatory checkpoint molecule selected from CD27, CD28,CD40, ICOS, OX40, GITR and CD137. In some embodiments, the immunecheckpoint molecule is an inhibitory checkpoint molecule selected fromA2AR, B7-H3, B7-H4, BTLA, CTLA-4, IDO, KIR, LAG3, PD-1, TIM3, and VISTA.In some embodiments, the compounds provided herein can be used incombination with one or more agents selected from KIR inhibitors, TIGITinhibitors, LAIR1 inhibitors, CD160 inhibitors, 2B4 inhibitors and TGFRbeta inhibitors.

In some embodiments, immune checkpoint inhibitors include inhibitorsagainst immune checkpoint molecules such as JAK1 and/or JAK2.

In some embodiments, immune checkpoint inhibitors include inhibitorsagainst immune checkpoint molecules such as CD96.

In some embodiments, the inhibitor of an immune checkpoint molecule isanti-PD1 antibody, anti-PD-L1 antibody, or anti-CTLA-4 antibody.

In some embodiments, the inhibitor of an immune checkpoint molecule isan inhibitor of PD-1, e.g., an anti-PD-1 monoclonal antibody. In someembodiments, the anti-PD-1 monoclonal antibody is nivolumab,pembrolizumab (also known as MK-3475), pidilizumab, SHR-1210, orAMP-224. In some embodiments, the anti-PD-1 monoclonal antibody isnivolumab or pembrolizumab. In some embodiments, the anti-PD1 antibodyis pembrolizumab.

In some embodiments, the inhibitor of an immune checkpoint molecule isan inhibitor of PD-L1, e.g., an anti-PD-L1 monoclonal antibody. In someembodiments, the anti-PD-L1 monoclonal antibody is BMS-935559, MED14736,MPDL3280A (also known as RG7446), or MSB0010718C. In some embodiments,the anti-PD-L1 monoclonal antibody is MPDL3280A or MED14736.

In some embodiments, the inhibitor of an immune checkpoint molecule isan inhibitor of CTLA-4, e.g., an anti-CTLA-4 antibody. In someembodiments, the anti-CTLA-4 antibody is ipilimumab.

In some embodiments, the inhibitor of an immune checkpoint molecule isan inhibitor of LAG3, e.g., an anti-LAG3 antibody. In some embodiments,the anti-LAG3 antibody is BMS-986016 or LAG525.

In some embodiments, the inhibitor of an immune checkpoint molecule isan inhibitor of GITR, e.g., an anti-GITR antibody. In some embodiments,the anti-GITR antibody is TRX518 or MK-4166. In some embodiments, theanti-GITR antibody is INCAGN01876.

In some embodiments, the inhibitor of an immune checkpoint molecule isan inhibitor of OX40, e.g., an anti-OX40 antibody or OX40L fusionprotein. In some embodiments, the anti-OX40 antibody is MEDI0562. Insome embodiments, the OX40L fusion protein is MEDI6383. In someembodiments, the anti-OX40 antibody is INCAGN01949.

In some embodiments, the inhibitor of an immune checkpoint molecule isan inhibitor of TIM3, e.g., an anti-TIM3 antibody.

In some embodiments Pim inhibitors of the invention can be combined withTIGIT inhibitors.

The Pim inhibitors of the present invention can be used in combinationwith one or more other anti-cancer agents including BET inhibitors(e.g., INCB054329, OTX015, and CPI-0610), LSD1 inhibitors (e.g.,GSK2979552 and INCB059872), HDAC inhibitors (e.g., panobinostat,vorinostat, and entinostat), DNA methyl transferase inhibitors (e.g.,azacitidine and decitabine), and other epigenetic modulators.

In some embodiments Pim inhibitors of the invention can be combined withBET inhibitors. In some embodiments Pim inhibitors of the invention canbe combined with LSD1 inhibitors. In some embodiments Pim inhibitors ofthe invention can be combined with HDAC inhibitors. In some embodimentsPim inhibitors of the invention can be combined with DNA methyltransferase inhibitors.

The Pim inhibitors of the present invention can be used in combinationwith one or more agents for the treatment of diseases such as cancer. Insome embodiments, the agent is an alkylating agent, a proteasomeinhibitor, a corticosteroid, or an immunomodulatory agent. Examples ofan alkylating agent include cyclophosphamide (CY), melphalan (MEL), andbendamustine. In some embodiments, the proteasome inhibitor iscarfilzomib. In some embodiments, the corticosteroid is dexamethasone(DEX). In some embodiments, the immunomodulatory agent is lenalidomide(LEN) or pomalidomide (POM).

One or more anti-inflammatory agents, steroids, immunosuppressants, ortherapeutic antibodies can also be combined with the salts of theinvention.

When more than one pharmaceutical agent is administered to a patient,they can be administered simultaneously, sequentially, or in combination(e.g., for more than two agents).

Formulation, Dosage Forms and Administration

The salts of the invention can be administered in the form of apharmaceutical composition. Thus the present disclosure provides acomposition comprising the salts of the invention, or any of theembodiments thereof, and at least one pharmaceutically acceptablecarrier. These compositions can be prepared in a manner well known inthe pharmaceutical art, and can be administered by a variety of routes,depending upon whether local or systemic treatment is indicated and uponthe area to be treated. Administration may be topical (includingtransdermal, epidermal, ophthalmic and to mucous membranes includingintranasal, vaginal and rectal delivery), pulmonary (e.g., by inhalationor insufflation of powders or aerosols, including by nebulizer;intratracheal or intranasal), oral or parenteral. Parenteraladministration includes intravenous, intraarterial, subcutaneous,intraperitoneal intramuscular or injection or infusion; or intracranial,e.g., intrathecal or intraventricular, administration. Parenteraladministration can be in the form of a single bolus dose, or may be,e.g., by a continuous perfusion pump. Pharmaceutical compositions andformulations for topical administration may include transdermal patches,ointments, lotions, creams, gels, drops, suppositories, sprays, liquidsand powders. Conventional pharmaceutical carriers, aqueous, powder oroily bases, thickeners and the like may be necessary or desirable.

This invention also includes pharmaceutical compositions which contain,as the active ingredient, a salt of the invention, in combination withone or more pharmaceutically acceptable carriers (excipients). In someembodiments, the composition is suitable for topical administration. Inmaking the compositions of the invention, the active ingredient istypically mixed with an excipient, diluted by an excipient or enclosedwithin such a carrier in the form of, e.g., a capsule, sachet, paper, orother container. When the excipient serves as a diluent, it can be asolid, semi-solid, or liquid material, which acts as a vehicle, carrieror medium for the active ingredient. Thus, the compositions can be inthe form of tablets, pills, powders, lozenges, sachets, cachets,elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solidor in a liquid medium), ointments containing, e.g., up to 10% by weightof the active compound, soft and hard gelatin capsules, suppositories,sterile injectable solutions and sterile packaged powders.

In preparing a formulation, the active compound can be milled to providethe appropriate particle size prior to combining with the otheringredients. If the active compound is substantially insoluble, it canbe milled to a particle size of less than 200 mesh. If the activecompound is substantially water soluble, the particle size can beadjusted by milling to provide a substantially uniform distribution inthe formulation, e.g., about 40 mesh.

The salts of the invention may be milled using known milling proceduressuch as wet milling to obtain a particle size appropriate for tabletformation and for other formulation types. Finely divided(nanoparticulate) preparations of the compounds of the invention can beprepared by processes known in the art see, e.g., WO 2002/000196.

Some examples of suitable excipients include lactose, dextrose, sucrose,sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates,tragacanth, gelatin, calcium silicate, microcrystalline cellulose,polyvinylpyrrolidone, cellulose, water, syrup and methyl cellulose. Theformulations can additionally include: lubricating agents such as talc,magnesium stearate and mineral oil; wetting agents; emulsifying andsuspending agents; preserving agents such as methyl- andpropylhydroxy-benzoates; and sweetening agents and flavoring agents. Thecompositions of the invention can be formulated so as to provide quick,sustained or delayed release of the active ingredient afteradministration to the patient by employing procedures known in the art.

In some embodiments, the pharmaceutical composition comprises silicifiedmicrocrystalline cellulose (SMCC) and at least one compound describedherein, or a pharmaceutically acceptable salt thereof. In someembodiments, the silicified microcrystalline cellulose comprises about98% microcrystalline cellulose and about 2% silicon dioxide w/w.

In some embodiments, the composition is a sustained release compositioncomprising at least a salt described herein, and at least onepharmaceutically acceptable carrier. In some embodiments, thecomposition comprises a salt described herein, and at least onecomponent selected from microcrystalline cellulose, lactose monohydrate,hydroxypropyl methylcellulose and polyethylene oxide. In someembodiments, the composition comprises at least one compound describedherein, or a pharmaceutically acceptable salt thereof, andmicrocrystalline cellulose, lactose monohydrate and hydroxypropylmethylcellulose. In some embodiments, the composition comprises at asalt described herein and microcrystalline cellulose, lactosemonohydrate and polyethylene oxide. In some embodiments, the compositionfurther comprises magnesium stearate or silicon dioxide. In someembodiments, the microcrystalline cellulose is Avicel PH102™. In someembodiments, the lactose monohydrate is Fast-flo 316™. In someembodiments, the hydroxypropyl methylcellulose is hydroxypropylmethylcellulose 2208 K4M (e.g., Methocel K4 M Premier™) and/orhydroxypropyl methylcellulose 2208 K100LV (e.g., Methocel KOOLV™). Insome embodiments, the polyethylene oxide is polyethylene oxide WSR 1105(e.g., Polyox WSR 1105™).

In some embodiments, a wet granulation process is used to produce thecomposition. In some embodiments, a dry granulation process is used toproduce the composition.

The compositions can be formulated in a unit dosage form, each dosagecontaining from about 5 to about 1,000 mg (1 g), more usually about 100mg to about 500 mg, of the active ingredient. In some embodiments, eachdosage contains about 10 mg of the active ingredient. In someembodiments, each dosage contains about 50 mg of the active ingredient.In some embodiments, each dosage contains about 25 mg of the activeingredient. The term “unit dosage forms” refers to physically discreteunits suitable as unitary dosages for human subjects and other mammals,each unit containing a predetermined quantity of active materialcalculated to produce the desired therapeutic effect, in associationwith a suitable pharmaceutical excipient. Solid oral dosage formsinclude, for example, tablets, capsules, and pills.

The components used to formulate the pharmaceutical compositions are ofhigh purity and are substantially free of potentially harmfulcontaminants (e.g., at least National Food grade, generally at leastanalytical grade, and more typically at least pharmaceutical grade).Particularly for human consumption, the composition is preferablymanufactured or formulated under Good Manufacturing Practice standardsas defined in the applicable regulations of the U.S. Food and DrugAdministration. For example, suitable formulations may be sterile and/orsubstantially isotonic and/or in full compliance with all GoodManufacturing Practice regulations of the U.S. Food and DrugAdministration.

The active compound may be effective over a wide dosage range and isgenerally administered in a therapeutically effective amount. It will beunderstood, however, that the amount of the compound actuallyadministered will usually be determined by a physician, according to therelevant circumstances, including the condition to be treated, thechosen route of administration, the actual compound administered, theage, weight and response of the individual patient, the severity of thepatient's symptoms and the like.

The therapeutic dosage of a compound of the present invention can varyaccording to, e.g., the particular use for which the treatment is made,the manner of administration of the compound, the health and conditionof the patient, and the judgment of the prescribing physician. Theproportion or concentration of a compound of the invention in apharmaceutical composition can vary depending upon a number of factorsincluding dosage, chemical characteristics (e.g., hydrophobicity), andthe route of administration. For example, the compounds of the inventioncan be provided in an aqueous physiological buffer solution containingabout 0.1 to about 10% w/v of the compound for parenteraladministration. Some typical dose ranges are from about 1 μg/kg to about1 g/kg of body weight per day. In some embodiments, the dose range isfrom about 0.01 mg/kg to about 100 mg/kg of body weight per day. Thedosage is likely to depend on such variables as the type and extent ofprogression of the disease or disorder, the overall health status of theparticular patient, the relative biological efficacy of the compoundselected, formulation of the excipient, and its route of administration.Effective doses can be extrapolated from dose-response curves derivedfrom in vitro or animal model test systems.

For preparing solid compositions such as for tablets, capsules, pills,or other oral dosage forms, the principal active ingredient is mixedwith a pharmaceutical excipient to form a solid preformulationcomposition containing a homogeneous mixture of a compound of thepresent invention. When referring to these preformulation compositionsas homogeneous, the active ingredient is typically dispersed evenlythroughout the composition so that the composition can be readilysubdivided into equally effective unit dosage forms such as tablets,pills and capsules. This solid preformulation is then subdivided intounit dosage forms of the type described above containing from, e.g.,about 0.1 to about 1000 mg of the active ingredient of the presentinvention.

The tablets or pills of the present invention can be coated or otherwisecompounded to provide a dosage form affording the advantage of prolongedaction. For example, the tablet or pill can comprise an inner dosage andan outer dosage component, the latter being in the form of an envelopeover the former. The two components can be separated by an enteric layerwhich serves to resist disintegration in the stomach and permit theinner component to pass intact into the duodenum or to be delayed inrelease. A variety of materials can be used for such enteric layers orcoatings, such materials including a number of polymeric acids andmixtures of polymeric acids with such materials as shellac, cetylalcohol and cellulose acetate.

The liquid forms in which the compounds and compositions of the presentinvention can be incorporated for administration orally or by injectioninclude aqueous solutions, suitably flavored syrups, aqueous or oilsuspensions, and flavored emulsions with edible oils such as cottonseedoil, sesame oil, coconut oil, or peanut oil, as well as elixirs andsimilar pharmaceutical vehicles.

Compositions for inhalation or insufflation include solutions andsuspensions in pharmaceutically acceptable, aqueous or organic solvents,or mixtures thereof, and powders. The liquid or solid compositions maycontain suitable pharmaceutically acceptable excipients as describedsupra. In some embodiments, the compositions are administered by theoral or nasal respiratory route for local or systemic effect.Compositions can be nebulized by use of inert gases. Nebulized solutionsmay be breathed directly from the nebulizing device or the nebulizingdevice can be attached to a face mask, tent, or intermittent positivepressure breathing machine. Solution, suspension, or powder compositionscan be administered orally or nasally from devices which deliver theformulation in an appropriate manner.

Topical formulations can contain one or more conventional carriers. Insome embodiments, ointments can contain water and one or morehydrophobic carriers selected from, e.g., liquid paraffin,polyoxyethylene alkyl ether, propylene glycol, white Vaseline®(petroleum jelly) and the like. Carrier compositions of creams can bebased on water in combination with glycerol and one or more othercomponents, e.g., glycerinemonostearate, PEG-glycerinemonostearate andcetylstearyl alcohol. Gels can be formulated using isopropyl alcohol andwater, suitably in combination with other components such as, e.g.,glycerol, hydroxyethyl cellulose and the like. In some embodiments,topical formulations contain at least about 0.1, at least about 0.25, atleast about 0.5, at least about 1, at least about 2, or at least about 5wt % of the compound of the invention. The topical formulations can besuitably packaged in tubes of, e.g., 100 g which are optionallyassociated with instructions for the treatment of the select indication,e.g., psoriasis or other skin condition.

The amount of compound or composition administered to a patient willvary depending upon what is being administered, the purpose of theadministration, such as prophylaxis or therapy, the state of thepatient, the manner of administration and the like. In therapeuticapplications, compositions can be administered to a patient alreadysuffering from a disease in an amount sufficient to cure or at leastpartially arrest the symptoms of the disease and its complications.Effective doses will depend on the disease condition being treated aswell as by the judgment of the attending clinician depending uponfactors such as the severity of the disease, the age, weight and generalcondition of the patient and the like.

The compositions administered to a patient can be in the form ofpharmaceutical compositions described above. These compositions can besterilized by conventional sterilization techniques, or may be sterilefiltered. Aqueous solutions can be packaged for use as is, orlyophilized, the lyophilized preparation being combined with a sterileaqueous carrier prior to administration. The pH of the compoundpreparations typically will be between 3 and 11, more preferably from 5to 9 and most preferably from 7 to 8. It will be understood that use ofcertain of the foregoing excipients, carriers or stabilizers will resultin the formation of pharmaceutical salts.

The therapeutic dosage of a compound of the present invention can varyaccording to, e.g., the particular use for which the treatment is made,the manner of administration of the compound, the health and conditionof the patient, and the judgment of the prescribing physician. Theproportion or concentration of a compound of the invention in apharmaceutical composition can vary depending upon a number of factorsincluding dosage, chemical characteristics (e.g., hydrophobicity), andthe route of administration. For example, the compounds of the inventioncan be provided in an aqueous physiological buffer solution containingabout 0.1 to about 10% w/v of the compound for parenteraladministration. Some typical dose ranges are from about 1 μg/kg to about1 g/kg of body weight per day. In some embodiments, the dose range isfrom about 0.01 mg/kg to about 100 mg/kg of body weight per day. Thedosage is likely to depend on such variables as the type and extent ofprogression of the disease or disorder, the overall health status of theparticular patient, the relative biological efficacy of the compoundselected, formulation of the excipient, and its route of administration.Effective doses can be extrapolated from dose-response curves derivedfrom in vitro or animal model test systems.

It is appreciated that certain features of the invention, which are, forclarity, described in the context of separate embodiments, can also beprovided in combination in a single embodiment (while the embodimentsare intended to be combined as if written in multiply dependent form).Conversely, various features of the invention which are, for brevity,described in the context of a single embodiment, can also be providedseparately or in any suitable subcombination.

The invention will be described in greater detail by way of specificexamples. The following examples are offered for illustrative purposes,and are not intended to limit the invention in any manner. Those ofskill in the art will readily recognize a variety of non-criticalparameters which can be changed or modified to yield essentially thesame results.

EXAMPLES Example 1 Asymmetric Synthesis ofN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamidePhosphoric Acid Salt

Step 1. Synthesis of tert-butyl[(3R,4R,5S)-4-{[tert-butyl(dimethyl)silyl]oxy}-1-((7R)-7-{[tert-butyl(dimethyl)silyl]oxy}-3-cyano-6,7-dihydro-5H-cyclopenta[b]pyridin-4-yl)-5-methylpiperidin-3-yl]carbamate(21)

To a stirred solution of(7R)-7-{[tert-butyl(dimethyl)silyl]oxy}-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carbonitrile(20) (see Example 5, 54.0 g, 173 mmol) (99.2% pure by HPLC) in anhydrousdimethyl sulfoxide (DMSO, 162 mL) in a 1-L RBF was added tert-butyl((3R,4R,5S)-4-{[tert-butyl(dimethyl)silyl]oxy}-5-methylpiperidin-3-yl)carbamatehydrochloride (9 (HCl)) (see U.S. Pat. Pub. No. 2014/0200227, para[0769], 70.0 g, 182.0 mmol, 1.05 equiv) and N,N-diisopropylethylamine(DIEA, 121 mL, 694 mmol, 4 equiv) at room temperature. The resultingreaction mixture was heated at 100° C. (oil bath) for 6 h. When LCMS andHPLC showed the reaction was complete (≥98.5% conversion), the reactionmixture was cooled to room temperature with a water bath, diluted withwater (400 mL), extracted with t-butylmethylether (TBME) twice (700 and400 mL). The organic layers were washed with brine (500 mL), dried overMgSO₄, filtered to remove the drying agent and concentrated in vacuo.The residue was purified by filtration chromatography (330 g silica gelcolumn; elution with 5% EtOAc/hexanes over 4 min, followed by 25%EtOAc/hexanes over 6 min to give the desired product, tert-butyl[(3R,4R,5S)-4-{[tert-butyl(dimethyl)silyl]oxy}-1-((7R)-7-{[tert-butyl(dimethyl)silyl]oxy}-3-cyano-6,7-dihydro-5H-cyclopenta[b]pyridin-4-yl)-5-methylpiperidin-3-yl]carbamate(21) (100.6 g, 98.1% pure by HPLC, 92% yield) as an off-white solid. ¹HNMR (400 MHz, DMSO-d₆) δ 8.48 (s, 1H), 6.75 (d, J=9.4 Hz, 1H), 5.01 (dd,J=6.9, 5.2 Hz, 1H), 3.61 (d, J=12.6 Hz, 1H), 3.52 (d, J=12.4 Hz, 1H),3.48-3.36 (m, 1H), 3.23 (t, J=9.5 Hz, 1H), 3.02 (t, J=11.6 Hz, 2H), 2.84(t, J=12.5 Hz, 1H), 2.74 (dt, J=15.2, 7.6 Hz, 1H), 2.32 (td, J=12.9, 8.0Hz, 1H), 1.82 (ddd, J=13.3, 8.1, 4.3 Hz, 1H), 1.75-1.60 (m, 1H), 1.36(s, 9H), 0.94 (d, J=6.5 Hz, 3H), 0.87 (s, 9H), 0.84 (s, 9H), 0.13 (s,3H), 0.08 (s, 6H), 0.06 (s, 3H) ppm; LCMS (EI) m/z 617.4 (C₃₂H₅₇N₄O₄Si₂,(M+H)⁺).

Step 2. Synthesis of tert-butyl((3R,4R,5S)-1-((7R)-3-(aminocarbonyl)-7-{[tert-butyl(dimethyl)silyl]oxy}-6,7-dihydro-5H-cyclopenta[b]pyridin-4-yl)-4-{[tert-butyl(dimethyl)silyl]oxy}-5-methylpiperidin-3-yl)carbamate (22)

To a stirred solution of tert-butyl[(3R,4R,5S)-4-{[tert-butyl(dimethyl)silyl]oxy}-1-((7R)-7-{[tert-butyl(dimethyl)silyl]oxy}-3-cyano-6,7-dihydro-5H-cyclopenta[b]pyridin-4-yl)-5-methylpiperidin-3-yl]carbamate(21) (65.0 g, 105 mmol) in ethanol (195 mL) in a 2-L RBF was added[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (Pd(dppf)₂C₂complexed with dichloromethane (1:1); 1.721 g, 2.107 mmol, 0.02 equiv),acetaldoxime (32.8 mL, 527 mmol, 5.0 equiv), and water (65 mL) at roomtemperature. The resulting reaction mixture was degassed and refilledwith N₂ three times before being heated to reflux (oil bath; temperatureat approximately 90° C.) for 6 h. An additional amount of acetaldoxime(32.8 mL, 527 mmol, 5.0 equiv) was added. The reaction mixture was thenheated to reflux (oil bath; temperature at approximately 90° C.) foranother 16 h. When LCMS and HPLC showed the reaction was complete(≥96.5% conversion), the hot reaction mixture was treated with water(390 mL) before being gradually cooled down to room temperature. Theresulting slurry was stirred at room temperature for an additional 30min before being collected by filtration. The wet cake was washed withwater (3×100 mL) before re-slurry in a mixture of acetonitrile and water(400 mL; acetonitrile to water: 1 to 3 by volume). The solids werecollected by filtration, washed with water (3×100 mL), and dried underhouse vacuum at room temperature overnight to afford the desiredproduct, tert-butyl((3R,4R,5S)-1-((7R)-3-(aminocarbonyl)-7-{[tert-butyl(dimethyl)silyl]oxy}-6,7-dihydro-5H-cyclopenta[b]pyridin-4-yl)-4-{[tert-butyl(dimethyl)silyl]oxy}-5-methylpiperidin-3-yl)carbamate(22), as a light brown powder, which was directly used in the subsequentreaction without further purification.

¹H NMR (400 MHz, DMSO-d₆) δ 8.16 (s, 1H), 7.84 (s, 1H), 7.46 (s, 1H),6.66 (d, J=9.6 Hz, 1H), 5.01 (dd, J=6.8, 4.7 Hz, 1H), 3.48-3.36 (m, 1H),3.23-3.07 (m, 3H), 3.01-2.88 (m, 1H), 2.82 (t, J=11.7 Hz, 1H), 2.74-2.58(m, 2H), 2.35 (dt, J=12.3, 6.0 Hz, 1H), 1.84 (dt, J=13.1, 6.6 Hz, 1H),1.72-1.56 (m, 1H), 1.35 (s, 9H), 0.90 (d, J=6.6 Hz, 3H), 0.87 (s, 9H),0.84 (s, 9H), 0.14 (s, 3H), 0.08 (s, 3H), 0.07 (s, 3H), 0.06 (s, 3H);LCMS (EI) m/z 635.4 (C₃₂H₅₉N₄O₅Si₂, (M+H)⁺).

Step 3. Synthesis of tert-butyl((3R,4R,5S)-1-(3-amino-7-{[tert-butyl(dimethyl)silyl]oxy}-6,7-dihydro-5H-cyclopenta[b]pyridin-4-yl)-4-{[tert-butyl(dimethyl)silyl]oxy}-5-methylpiperidin-3-yl)carbamate(23)

To a stirred solution of tert-butyl((3R,4R,5S)-1-((7R)-3-(aminocarbonyl)-7-{[tert-butyl(dimethyl)silyl]oxy}-6,7-dihydro-5H-cyclopenta[b]pyridin-4-yl)-4-{[tert-butyl(dimethyl)silyl]oxy}-5-methylpiperidin-3-yl)carbamate(22) (66.9 g, 105 mmol) in tetrahydrofuran (THF, 470.0 mL) in a 2-L 3neck RBF at 0° C. was added tetra-N-butylammonium bromide (34.30 g,105.4 mmol, 1.0 equiv) and a solution of 3.0 M sodium hydroxide in water(158.0 mL, 474.1 mmol, 4.5 equiv). 1,3-Dibromo-5,5-dimethylhydantoin(21.52 g, 73.75 mmol, 0.7 equiv) was then added portion-wise over 44 minto control the reaction temperature at 5 to 10° C. The resulting darkbrown solution was stirred at 5 to 10° C. for an additional 20 min. WhenLCMS showed the reaction was complete, the reaction mixture was dilutedwith MTBE (100 mL), quenched with a 10% aqueous solution of Na₂S₂O₃ (300mL) and water (200 mL). The two phases were separated, and the aqueousphase was extracted with MTBE (2×600 mL). The combined organic layerswere washed with water (500 mL), dried over MgSO₄, filtered to removethe drying agent and concentrated in vacuo to afford the crude productas a brown foamy solid (63.7 g). The crude product was purified byfiltration chromatography (330 g silica gel column; elution with 20%EtOAc/hexanes for 6 min, followed by 45% EtOAc/hexanes for 8 min,followed by recrystallization from heptane (180 mL) to afford thedesired product, tert-butyl((3R,4R,5S)-1-(3-amino-7-{[tert-butyl(dimethyl)silyl]oxy}-6,7-dihydro-5H-cyclopenta[b]pyridin-4-yl)-4-{[tert-butyl(dimethyl)silyl]oxy}-5-methylpiperidin-3-yl)carbamate (23) (46.2 g, 100.0% pure byHPLC, 72% yield for two steps) as an off-white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 7.80 (s, 1H), 6.61 (d, J=9.7 Hz, 1H), 4.88(dd, J=6.8, 3.6 Hz, 1H), 4.81 (s, 2H), 3.52 (td, J=9.8, 4.6 Hz, 1H),3.14 (t, J=9.5 Hz, 1H), 3.04-2.84 (m, 3H), 2.85-2.68 (m, 2H), 2.48-2.42(m, 1H), 2.23 (dq, J=13.2, 6.9 Hz, 1H), 1.79 (dq, J=12.6, 4.4 Hz, 2H),1.35 (s, 9H), 0.89 (d, J=6.6 Hz, 3H), 0.85 (s, 18H), 0.10 (s, 3H), 0.08(s, 3H), 0.06 (s, 3H), −0.00 (s, 3H); LCMS (EI) m/z 607.4(C₃₁H₅₉N₄O₄Si₂, (M+H)⁺).

Step 4. Synthesis of tert-butyl{(3R,4R,5S)-4-{[tert-butyl(dimethyl)silyl]oxy}-1-[(7R)-7-{[tert-butyl(dimethyl)silyl]oxy}-3-({[6-(2,6-difluorophenyl)-5-fluoropyridin-2-yl]carbonyl}amino)-6,7-dihydro-5H-cyclopenta[b]pyridin-4-yl]-5-methylpiperidin-3-yl}carbamate(25)

To a stirred solution of tert-butyl((3R,4R,5S)-1-(3-amino-7-{[tert-butyl(dimethyl)silyl]oxy}-6,7-dihydro-5H-cyclopenta[b]pyridin-4-yl)-4-{[tert-butyl(dimethyl)silyl]oxy}-5-methylpiperidin-3-yl)carbamate(23) (100.0 g, 164.8 mmol) and6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxylic acid (24) (see U.S.Pat. Pub. No. 2014/0200227, para [0625] 50.05 g, 197.7 mmol, 1.2 equiv)in anhydrous N,N-dimethylformamide (DMF, 320.0 mL) was addedN,N-diisopropylethylamine (DIEA, 137.8 mL, 790.8 mmol, 4.8 equiv) atroom temperature. After stirring at room temperature for 10 min, thereaction mixture was treated withN,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uroniumhexafluorophosphate (HATU, 150.3 g, 395.4 mmol, 2.4 equiv) portion-wiseto control the temperature at 15 to 30° C. The reaction mixture was thenstirred at room temperature for 30 min. When LCMS and HPLC showed thereaction was complete, the reaction mixture was filtered into water(1300 mL) with stirring. The resulting slurry was stirred at roomtemperature for an additional 30 min. The solids were collected byfiltration and washed with a mixture of 50% acetonitrile and water (50%by volume; 2×200 mL). The wet solid was then treated with acetonitrile(420 mL) and the resulting slurry was heated at 70° C. until a clearsolution is generated. Water (320 mL) was added slowly to the solutionat 70° C. and the resulting mixture was gradually cooled to roomtemperature and stirred at room temperature for 30 min. The solids werecollected by filtration, washed with a mixture of acetonitrile and water(50% by volume; 320 mL) and dried in a vacuum oven at 50° C. forovernight to afford the desired product, tert-butyl{(3R,4R,5S)-4-{[tert-butyl(dimethyl)silyl]oxy}-1-[(7R)-7-{[tert-butyl(dimethyl)silyl]oxy}-3-({[6-(2,6-difluorophenyl)-5-fluoropyridin-2-yl]carbonyl}amino)-6,7-dihydro-5H-cyclopenta[b]pyridin-4-yl]-5-methylpiperidin-3-yl}carbamate(25) (131.8 g, 99.7% pure by HPLC, 95% yield) as white powder.

¹H NMR (400 MHz, DMSO-d₆) δ 10.39 (s, 1H), 8.95 (s, 1H), 8.36 (dd,J=8.7, 4.0 Hz, 1H), 8.19 (t, J=8.9 Hz, 1H), 7.67-7.55 (m, 1H), 7.27 (t,J=8.5 Hz, 3H), 6.46 (d, J=9.6 Hz, 1H), 5.00 (dd, J=6.8, 4.2 Hz, 1H),3.40 (dd, J=9.3, 5.9 Hz, 1H), 3.05 (t, J=9.4 Hz, 2H), 2.93 (t, J=10.8Hz, 4H), 2.82 (dt, J=15.8, 8.1 Hz, 1H), 2.56 (t, J=11.7 Hz, 1H),2.40-2.26 (m, 1H), 1.92-1.79 (m, 1H), 1.54 (s, OH), 1.32 (s, 9H), 0.87(s, 11H), 0.84 (s, 9H), 0.71 (d, J=6.5 Hz, 4H), 0.14 (s, 4H), 0.07 (s,3H), 0.04 (s, 4H), 0.01 (s, 3H); LCMS (EI) m/z 842.4 (C₄₃H₆₃F₃N₅O₅Si₂,(M+H)⁺).

Step 5. Synthesis ofN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamideDihydrochloric Acid (Compound 1 Dihydrochloric Acid Salt)

A solution of 4.0 M hydrogen chloride in 1,4-dioxane (13.9 L, 55600mmol, 15.0 equiv) was added into a slurry of tert-butyl{(3R,4R,5S)-4-{[tert-butyl(dimethyl)silyl]oxy}-1-[(7R)-7-{[tert-butyl(dimethyl)silyl]oxy}-3-({[6-(2,6-difluorophenyl)-5-fluoropyridin-2-yl]carbonyl}amino)-6,7-dihydro-5H-cyclopenta[b]pyridin-4-yl]-5-methylpiperidin-3-yl}carbamate(25) (3121.0 g, 3706.0 mmol) in methanol (19.5 L) at room temperature.The internal temperature rose from 17.3 to 36.8° C. during addition ofthe HCL solution in 1,4-dioxane. The resulting light yellow solution wasstirred at room temperature for 22 h and solids (2 HCl salt) started toprecipitate out of the solution within 2 h. When LCMS and HPLC indicatedthe reaction was complete, the solids were collected by filtration,washed with heptane (5 L), and dried on the filter under vacuumovernight to afford the desired product,N-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamidedihydrochloride (Compound 1 dihydrochloric acid salt) (2150.0 g, 99.1%pure by HPLC, 98% yield) as white powder. ¹H NMR (400 MHz, DMSO-d₆) δ10.57 (s, 1H), 8.57 (s, 3H), 8.36 (dd, J=8.7, 4.1 Hz, 1H), 8.22 (t,J=8.8 Hz, 1H), 7.69 (ddd, J=15.1, 8.5, 6.7 Hz, 1H), 7.34 (t, J=8.1 Hz,2H), 5.27 (t, J=7.0 Hz, 1H), 4.02 (d, J=11.1 Hz, 1H), 3.44 (d, J=11.3Hz, 1H), 3.29 (t, J=12.2 Hz, 1H), 3.13 (q, J=8.5, 7.4 Hz, 2H), 3.09-2.97(m, 2H), 2.77 (t, J=12.3 Hz, 1H), 2.56-2.50 (m, 1H), 1.95 (dq, J=14.9,7.7, 7.2 Hz, 1H), 1.66-1.50 (m, 1H), 0.70 (d, J=6.5 Hz, 3H); LCMS (EI)m/z 514.2 (C₂₆H₂₇F₃N₅O₃, (M+H)⁺).

Step 6. Synthesis ofN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1 Free Base)

A 200 L glass reactor was assembled with overhead stirring, condenser,thermocouple, addition funnel, and a nitrogen inlet and the apparatuswas purged with nitrogen. Sodium carbonate (18,264 g) and potable water(86.2 L) were charged to the reactor and stirred for about 50 minutesuntil a solution was obtained. Methylene chloride (107.8 L) was chargedto the reactor.N-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamidedihydrochloride (Compound 1 dihydrochloric acid salt) (4300 g) wascharged over about 27 minutes to the reactor while maintaining thetemperature at about 27° C. and the reaction mixture was stirred atabout 23° C. for about 22 hours until a clear solution was obtained. Thephases were separated, and the aqueous phase was extracted withmethylene chloride (26.9 L). The combined organic phases were washedtwice with potable water (26.9 L per wash) and concentrated underreduced pressure at about 54° C. to afford the desired product,N-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1 free base, 3838 g), as a light yellow solid, which wasdirectly used in the subsequent reaction without further purification.LCMS (EI) m/z 514.2 (C₂₆H₂₇F₃N₅O₃, (M+H)⁺).

Step 7. Synthesis ofN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamidePhosphoric Acid (Compound 1 Phosphoric Acid Salt)

A 100 L glass reactor was assembled with overhead stirring, condenser,thermocouple, addition funnel, and a nitrogen inlet and the apparatuswas purged with nitrogen. Separately, a phosphoric acid solution wasprepared by thoroughly mixing an aqueous solution of 85% H₃PO₄ (980 g)and 2-propanol (IPA, 5.7 L) at room temperature. The phosphoric acidsolution was polish filtered through an in-line filter. Separately,methanol (19.0 L), USP purified water (1.9 L), and 2-propanol werepolish filtered through an in-line filter. Filtered methanol (19.0 L),N-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide(Compound 1 free base) (3800 g), filtered USP purified water (1.9 L),and filtered 2-propanol (15.2 L) were charged sequentially to thereactor. The reaction mixture was heated to about 56° C. Filteredphosphoric acid solution (5.8 L) was charged to the reactor over about45 minutes while maintaining the temperature at about 59° C. Thecontainer was rinsed into the reaction mixture with filtered 2-propanol(5.8 L) while maintaining the temperature at about 62° C. Filtered2-propanol (15.2 L) was charged while maintaining the temperature atabout 58° C., and the reaction mixture was stirred at about 57° C. forabout 1.5 hours. The reaction mixture was cooled to about 26° C. andstirred at about 17° C. for about 3.5 hours. The reaction mixture wasfiltered and the filter cake was washed sequentially with filtered2-propanol (22.8 L) and filtered heptane (prepared separately by polishfiltering 22.8 L of heptane through an in-line filter). The product wasdried on the filter and then dried under reduced pressure at 20-54° C.to afford the desired product,N-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamidephosphoric acid salt (Compound 1 phosphoric acid salt) (3952 g, >99.0%pure by HPLC, 87.3% yield) as a white to off-white crystalline solid.

¹H NMR (400 MHz, DMSO-d₆) δ 10.43 (s, 1H), 9.25 (s, 1H), 8.38 (dd,J=8.7, 4.1 Hz, 1H), 8.20 (t, J=8.8 Hz, 1H), 7.66 (ddd, J=15.2, 8.4, 6.7Hz, 1H), 7.32 (t, J=8.6 Hz, 2H), 7.19 (s, 2H), 4.82 (dd, J=6.9, 4.5 Hz,1H), 3.22 (d, J=6.9 Hz, 1H), 3.18-2.99 (m, 3H), 2.87 (d, J=8.0 Hz, 2H),2.79-2.67 (m, 1H), 2.59 (t, J=11.5 Hz, 1H), 2.27 (dt, J=13.5, 6.6 Hz,1H), 1.92-1.74 (m, 1H), 1.58-1.39 (m, 1H), 0.70 (d, J=6.5 Hz, 3H) ppm;¹³C NMR (100 MHz, DMSO-d₆) δ 163.1, 160.5, 159.5 (dd, J_(CF)=249.4, 6.6Hz), 159.0 (d, J_(CF)=261.8 Hz), 146.0 (d, J_(CF)=4.2), 144.6, 136.1(J_(CF)=18.6), 140.8, 132.8 (t, J_(CF)=10.6 Hz), 130.6, 128.5, 126.2 (d,J_(CF)=20.5), 125.4 (d, J_(CF)=6.4), 112.4 (d, J_(CF)=20.8 Hz), 110.7(td, J_(CF)=19.5, 3.6 Hz), 74.9, 72.8, 55.7, 54.0, 52.4, 37.7, 33.1,26.9, 14.4 ppm; ¹⁹F NMR (376 MHz, DMSO-d₆) δ −117.26 (m, 1F), −113.77(m, 2F) ppm; LCMS (EI) m/z 514.2 (C₂₆H₂₇F₃N₅O₃, (M+H)⁺).

Results of quantitative elemental microanalysis for carbon, hydrogen,and nitrogen on Compound 1 phosphoric acid salt are in agreement withthe proposed empirical formula (C₂₆H₂₉F₃N₅O₇P). Anal. Calcd forC₂₆H₂₉F₃N₅O₇P: C, 51.07; H, 4.78; N, 11.45. Found: C, 51.16; H, 4.70; N,11.56.

Example 2 X-Ray Powder Diffraction (XRPD) of Compound 1 Phosphoric AcidSalt Form I

Form I of crystallineN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamidephosphoric acid salt, (see Example 1, Step 7) was characterized by XRPD.The X-Ray Power Diffraction (XRPD) was obtained from Rigaku MiniFlexX-ray Powder Diffractometer (XRPD). The general experimental proceduresfor XRPD were: (1) X-ray radiation from copper at 1.054056 Å with K_(β)filter; (2) X-ray power at 30 KV, 15 mA; and (3) the sample powder wasdispersed on a zero-background sample holder. The general measurementconditions for XRPD were: Start Angle 3 degrees; Stop Angle 45 degrees;Sampling 0.02 degrees; and Scan speed 2 degree/min. The XRPD pattern isshown in FIG. 1 and XRPD data is provided in Table 1.

TABLE 1 XRPD Data Form I 2-Theta (°) Height H % 4.6 1078 100 8.7 52 4.89.4 377 35 12.5 160 14.8 13.1 926 85.9 14.0 62 5.8 16.2 937 86.9 17.4534 49.6 17.9 717 66.5 18.8 708 65.7 19.4 616 57.1 20.3 199 18.5 21.1960 89.1 22.3 294 27.3 23.0 700 65 24.8 746 69.3 25.2 315 29.2 25.8 10910.1 26.4 364 33.8 27.6 165 15.3 28.9 84 7.8 29.4 92 8.6 30.2 136 12.630.7 77 7.2 33.3 99 9.2 34.1 165 15.3 34.9 168 15.5 35.5 199 18.5 36.056 5.2 37.0 49 4.5 37.6 64 5.9 38.2 246 22.8 38.6 80 7.4 39.8 67 6.340.1 64 5.9 40.9 66 6.2 41.7 119 11 42.1 84 7.8 43.7 82 7.6 43.9 43 4

Example 3 Differential Scanning Calorimetry (DSC) of Compound 1Phosphoric Acid Salt (Form I)

The crystalline solid,N-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamidephosphoric acid salt, (see Example 1, Step 7) was characterized by DSC.The DSC was obtained from TA Instruments Differential ScanningCalorimetry, Model Q200 with autosampler. The DSC instrument conditionswere as follows: 30-350° C. at 10° C./min; Tzero aluminum sample pan andlid; and nitrogen gas flow at 50 mL/min. The DSC thermogram is shown inFIG. 2. The DSC thermogram revealed a major endothermal event (believedto be a melting/decomposition) at an onset temperature of 238.8° C. witha peak temperature of 247.1° C. and a small endothermal event at anonset temperature of 193.1° C. with a peak temperature at 198.4° C.Multiple lots of Form I were characterized by DSC, each having athermogram with a major endothermal peak occurring within the range of249.7 to 254.7° C.

Example 4 Thermogravimetric Analysis (TGA) of Compound 1 Phosphoric AcidSalt (Form I)

The crystalline solid,N-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamidephosphoric acid salt, (see Example 1, Step 7) was characterized by TGA.The TGA was obtained from TA Instrument Thermogravimetric Analyzer,Model Q500. The general experimental conditions for TGA were: ramp from20° C. to 600° C. at 20° C./min; nitrogen purge, gas flow at 40 mL/minfollowed by balance of the purge flow; sample purge flow at 60 mL/min;platinum sample pan. The TGA thermogram is shown in FIG. 3. A weightloss of about 2% below 200° C. was observed and believed to beassociated with loss of moisture and residual solvents. Karl-Fischeranalysis of various synthetic lots of Form I was conducted, eachexperiment revealing a water content within the range of 1.40-1.50%indicating that Form I may be a hydrate, such as a hemihydrate.

Example 5 Asymmetric Synthesis of(7R)-7-{[tert-Butyl(dimethyl)silyl]oxy}-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carbonitrile(20)

Step 1. 6,7-Dihydro-5H-cyclopenta[b]pyridine 1-oxide (11)

To a mixture of 6,7-dihydro-5H-cyclopenta[b]pyridine (10) (10.0 g, 83.9mmol) in methanol (50 mL) was added urea hydrogen peroxide adduct (UHP,24 g, 250 mmol, 3.0 equiv.) and methyltrioxorhenium(VII) (80 mg, 0.3mmol, 0.0036 equiv.) at room temperature. The resulting mixture wasstirred at room temperature overnight before being concentrated underreduced pressure to remove methanol. After no more distillate wasobserved, methylene chloride (100 mL) was added and the concentrationwas continued. The resulting residue was treated with methylene chloride(100 mL) and stirred at room temperature for 10 minutes. The solids werefiltered and extracted with methylene chloride for 3 times (3×50 mL).The combined filtrates were dried with anhydrous sodium sulfate (Na₂SO₄)and sodium bisulfite (NaHSO₃) before being filtered through a silica gelpad (SiO₂). The pad was rinsed with 10% MeOH in CH₂CH₂ (100 mL) for 3times. The combined filtrates were concentrated to give the desiredproduct, 6,7-dihydro-5H-cyclopenta[b]pyridine 1-oxide (11, 11.5 g, 99%yield) as off-white solid, which was used in the subsequent reactionwithout further purification. For 11: ¹HNMR (300 MHz, CDCl₃) δ 8.05 (d,J=6.0 Hz, 1H), 7.08 (m, 2H), 3.15. (t, J=6.0 Hz, 2H), 3.00. (t, J=6.0Hz, 2H), 2.16 (m, 2H) ppm; LCMS (EI) m/z 136 (C₈H₉NO, (M+H)⁺).

Step 2. 4-Chloro-6,7-dihydro-5H-cyclopenta[b]pyridine (12)

6,7-Dihydro-5H-cyclopenta[b]pyridine 1-oxide (11, 12.0 g, 84.3 mmol) wasslowly added into phosphoryl chloride (POCl₃, 24 mL, 250 mmol, 3.0equiv) and toluene (48 mL) at 80-85° C. (internal temperature) withabout 0.2 g per portion over 2 hours. After the completion of addition,the reaction mixture was continued to stir at 80-85° C. (internaltemperature) for 3 hours. The excess amount of POCl₃ and toluene wasremoved under reduced pressure at 60° C. The resulting residue was thenpoured to a cooled mixture of ice (50 g) and the saturated aqueoussodium carbonate solution (Na₂CO₃, 50 mL). The pH of the mixture wasadjusted to 8 using 25% aqueous sodium hydroxide solution (NaOH, about30 mL). The mixture was then extracted with methylene chloride (3×100mL). The combined organic extracts were dried over anhydrous Na₂SO₄ andfiltered through a pad of silica gel (SiO₂, 36 g) and charcoal (6 g, onthe top of silica gel). The pad was rinsed with ethyl acetate (4×100 mL)until no desired product came out from the silica gel pad. The combinedfiltrates were then concentrated under reduced pressure to give thecrude desired product (12, 10.93 g). To the mixture of crude4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine (12, 10.7 g, 69.6 mmol) inTHF (50 mL) was added 3-chlorobenzoic acid (11 g, 73 mmol) at roomtemperature. The mixture was then stirred at room temperature until3-chlorobenzoic acid was dissolved. The mixture was concentrated underreduced pressure to generate the crude salt as solids, which was treatedwith hexanes (32 mL). The resulting suspension was stirred at roomtemperature for 10 minutes. The solids were collected by filtration,washed with hexanes (2×32 mL). More solids were collected from thefiltrates. The combined wet solids (about 18 g) were dissolved inmethylene chloride (100 mL) and treated with the saturated aqueousNa₂CO₃ solution (2×20 mL). The aqueous layers were back extracted withmethylene chloride (50 mL). The combined organic extracts were driedwith anhydrous Na₂SO₄, filtered and concentrated to give the purifieddesired product, 4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine (12, 7.1g), as dark oil. For 12: ¹HNMR (300 MHz, CDCl₃) δ 8.20 (d, J=3.0 Hz,1H), 7.02 (d, J=3.0 Hz, 1H), 3.06. (t, J=6.0 Hz, 2H), 2.98. (t, J=6.0Hz, 2H), 2.12 (m, 2H) ppm; LCMS (EI) m/z 154/156 (C₈H₈ClN, (M+H)⁺).

Step 3. 4-Chloro-6,7-dihydro-5H-cyclopenta[b]pyridine 1-oxide (13)

To a mixture of 4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine (12, 100.0g, 618.4 mmol) in methanol (500 mL) was added urea hydrogen peroxideadduct (UHP, 1.5 eq, 87.3 g, 928 mmol, 1.5 equiv) andmethyltrioxorhenium(VII) (925 mg, 3.71 mmol, 0.006 equiv) at roomtemperature. The resulting mixture was stirred at room temperature for 3hours. The reaction mixture was concentrated under reduced pressure toremove methanol. Methylene chloride (500 mL) was added and theconcentration was continued. The resulting residue was treated withmethylene chloride (500 mL) and the resulting mixture was stirred for 30minutes. The solids were filtered and extracted with methylene chloridefor (4×500 mL). The combined filtrates were dried with anhydrous sodiumsulfate (Na₂SO₄) and sodium bisulfite (NaHSO₃). The mixture was thenfiltered through a pad of silica gel (SiO₂, 180 g) and the activatedcharcoal (10 g). The pad was rinsed with 10% MeOH in CH₂CH₂ (500 mL) 3times. The combined filtrates were concentrated under reduced pressureto give the desired product,4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine 1-oxide (13, 104 g), asoff-white solids. For 13: ¹HNMR (300 MHz, DMSO-d₆) δ 8.04 (d, J=6.0 Hz,1H), 7.34 (d, J=6.0 Hz, 1H), 2.98. (m, 4H), 2.08 (m, 2H) ppm; LCMS (EI)m/z 170/172 (C₈H₈ClNO, (M+H)⁺).

Step 4. 4-Chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-7-yl acetate (14)

4-Chloro-6,7-dihydro-5H-cyclopenta[b]pyridine 1-oxide (13) (65.0 g, 340mmol) was added to the mixture of acetic anhydride (Ac₂O, 98 mL, 1000mmol) and toluene (325 mL) in portions (6 g per portion) at 80-85° C.over 1 hour. After the addition was complete, the resulting mixture wasstirred at 80-85° C. for 3 hours. When the reaction completion wasindicated by LCMS and/or HPLC, the reaction mixture was concentratedunder reduced pressure to remove acetic anhydride and toluene. The crudedesired product, 4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-7-ylacetate (14), obtained was used directly in the subsequent reactionwithout further purification. For 14: ¹HNMR (300 MHz, DMSO-d₆) δ 8.38(d, J=6.0 Hz, 1H), 7.21 (d, J=6.0 Hz, 1H), 6.14. (m, 1H), 3.08 (m, 1H),2.93 (m, 1H), 2.64 (m, 1H), 2.10 (s, 3H), 2.07 (m, 1H) ppm; LCMS (EI)m/z 212/214 (C₁₀H₁₀ClNO₂, (M+H)⁺).

Step 5. 4-Chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-7-ol (15)

To a mixture of 4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-7-ylacetate (14, 73 g, 340 mmol, telescoped from the last step) in MeOH (130mL) and water (130 mL) was added potassium carbonate (K₂CO₃, 130 g, 920mmol) at 0° C. The resulting mixture was gradually warmed to roomtemperature and stirred at room temperature for 4 hours. The reactionmixture was then concentrated under reduced pressure to remove methanol.The precipitated solids were collected by filtration, washed with water(3×100 mL), and dried to give the first portion of the crude product(15). The aqueous layers were extracted with DCM (3×300 mL). Thecombined organic extracts were dried over anhydrous Na₂SO₄, filtered andconcentrated to give the second portion of the crude product (15). Thecrude desired product, 4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-7-ol(15, 61.9 g, 93% yield for 2 steps) obtained from two portions as darkbrown oil, was directly used in the subsequent reaction without furtherpurification. For 15: ¹HNMR (300 MHz, DMSO-d₆) δ 8.34 (d, J=3.0 Hz, 1H),7.18 (d, J=3.0 Hz, 1H), 5.26. (t, J=3.0 Hz, 1H), 3.08 (m, 1H), 2.86 (m,1H), 2.60 (m, 1H), 2.09 (m, 1H) ppm; LCMS (EI) m/z 170/172 (C₈H₈ClNO,(M+H)⁺).

Step 6. 4-Chloro-5H-cyclopenta[b]pyridin-7(6H)-one (16)

To the solution of 4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-7-ol(15, 110.3 g, 650.32 mmol) in methylene chloride (800 mL) was addedN,N-diisopropylethylamine (339.5 g, 2627.0 mmol) at room temperature.After the mixture was cooled to 0° C., pyridine-sulfur trioxide (1:1)(200.0 g, 1256.6 mmol) in dimethyl sulfoxide (DMSO, 800 g) was addeddropwise over 35 minutes and the resulting mixture was stirred at 0° C.for 1 hour. The reaction progress was monitored by HPLC and about 30% ofstarting material 15 was left. Additional amounts of pyridine-sulfurtrioxide (1:1) (138 g, 866.8 mmol) and N,N-diisopropylethylamine (79.8g, 617 mmols) were added, and the reaction mixture was stirred under anice bath for another 2 hours. Water (1000 mL) was added to the reactionmixture, and the resulting mixture was concentrated under reducedpressure to remove methylene chloride. The resulting residue wascarefully poured into saturated aqueous NaHCO₃ (2000 mL). The solidswere collected by filtration, washed with water (2×200 mL), and dried togive the first portion of the crude desired product (16). The aqueousfiltrate was extracted with methylene chloride (2×200 mL). The combinedorganic extracts were dried over anhydrous Na₂SO₄, filtered andconcentrated to give the second portion of the crude desired product(16). The combined crude desired product was purified by silica gelflash chromatography (SiO₂, eluted with 0 to 50% ethyl acetate/hexane)to provide the desired product,4-chloro-5H-cyclopenta[b]pyridin-7(6H)-one (16, 87 g, 80% yield), asyellow to brown oil, which solidified upon standing in vacuum. For 16:¹HNMR (300 MHz, DMSO-d₆) δ 8.68 (d, J=6.0 Hz, 1H), 7.48 (d, J=6.0 Hz,1H), 3.18 (m, 2H), 2.81 (m, 2H) ppm; LCMS (EI) m/z 168/170 (C₈H₆ClNO,(M+H)⁺).

Step 7. (R)-4-Chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-7-ol (17)

To the solution of 4-chloro-5H-cyclopenta[b]pyridin-7(6H)-one (16, 138.0g, 823.4 mmol) and triethylamine (TEA, 573.8 mL, 4117 mmol) in methylenechloride (1100 mL) was added RuCl(p-cymene)[(R,R)-Ts-DPEN] (1.31 g, 2.06mmol) at room temperature. The resulting mixture was degased, cooleddown to 5-10° C., and stirred at 5-10° C. under nitrogen. Formic acid(155.3 mL, 4117 mmol) was then added slowly (internal temperature) tothe reaction mixture at 7-14° C. After the addition, the reactionmixture gradually warmed to room temperature and stirred at roomtemperature for 22 hours. When LC/MS and HPLC showed the reaction wascomplete, the reaction mixture was quenched with the saturated aqueoussodium bicarbonate (NaHCO₃) solution (1000 mL) and water (1500 mL). Thetwo layers were separated, and the aqueous layer was extracted withmethylene chloride (3×500 mL). The organic layers were combined, driedover anhydrous Na₂SO₄, filtered and concentrated to give the crudedesired product, (R)-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-7-ol(17, assumed 100% yield) as yellow to brown oil, which was directly usedfor the subsequent reaction without further purification. For 17: ¹HNMR(300 MHz, DMSO-d₆) δ 8.33 (d, J=6.0 Hz, 1H), 7.19 (d, J=6.0 Hz, 1H),5.27 (m, 1H), 3.08 (m, 1H), 2.86 (m, 1H), 2.57 (m, 1H), 2.10 (m, 1H)ppm; LCMS (EI) m/z 170/172 (C₈H₈ClNO, (M+H)⁺).

Step 8.(R)-7-(tert-Butyldimethylsilyloxy)-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine(18)

To the stirred solution of(R)-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridin-7-ol (17, 172.25 g,1015.6 mmol) in anhydrous methylene chloride (2500 mL) was addedtert-butyldimethylsilyl chloride (172.25 g, 1015.6 mmol) and1H-imidazole (101.3 g, 1472 mmol) at 0-5° C. The resulting reactionmixture was stirred at room temperature for overnight. When LC/MS andHPLC showed the reaction was complete, the reaction mixture was quenchedwith water (1000 mL). The two layers were separated, and the aqueouslayer was extracted with methylene chloride (3×500 mL). The combinedorganic layers were dried over anhydrous Na₂SO₄, filtered andconcentrated to give the crude desired product (18). The purification ofthe crude product on silica gel chromatography (SiO₂, eluting with 0-10%ethyl acetate in hexanes) provided the desired product,(R)-7-(tert-butyldimethylsilyloxy)-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine(18, 253.0 g, 87.8% yield for 2 steps) as yellow to brown oil. For 18:¹HNMR (300 MHz, DMSO-d₆) δ 8.36 (d, J=3.0 Hz, 1H), 7.13 (d, J=3.0 Hz,1H), 5.21 (m, 1H), 3.08 (m, 1H), 2.81 (m, 1H), 2.43 (m, 1H), 2.05 (m,1H), 0.93 (s, 9H), 0.22 (s, 3H), 0.16 (s, 3H) ppm; LCMS (EI) m/z 284/286(C₁₄H₂₂ClNOSi, (M+H)⁺).

Step 9.(R)-7-(tert-Butyldimethylsilyloxy)-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carbaldehyde(19)

To the stirred solution of 2,2,6,6-tetramethyl-piperidine (63.0 g, 446mmol) in anhydrous tetrahydrofuran (THF, 680 mL) at −24° C. to −50° C.was added a solution of 2.5 M n-butyl lithium in hexane (180 mL, 450mmol). The resulting mixture was warmed up to >−32° C. and stirred at−32° C. to 0° C. for 30 minutes and then cooled to −78° C. (externalbath).(R)-7-(tert-butyldimethylsilyloxy)-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine(18, 106.0 g, 373 mmol) in anhydrous tetrahydrofuran (380 mL) was addeddropwise to the above cooled mixture at −74° C. to −76° C. (internaltemperature). After the resulting dark brown solution was stirred at−74° C. (internal temperature) for 90 minutes, anhydrousN,N-dimethylformamide (DMF, 130 g, 1129 mmol) was added and the internaltemperature was maintained at −76° C. to −70° C. during addition. Theresulting reaction mixture was continued to stir at −74° C. to −72° C.for 2 hours before being quenched with 1 N aqueous HCl solution (500 mL)and water (500 mL). The two phases were separated, and the aqueous phasewas extracted with MTBE (2×250 mL). The combined organic phases werewashed with brine (2×250 mL) and concentrated under reduced pressure togive the crude product (19). The purification of the crude product onsilica gel chromatography (SiO₂, eluting with 0-8% ethyl acetate inhexane) provided the desired product,(R)-7-(tert-butyldimethylsilyloxy)-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carbaldehyde(19, 110.7 g, 94% yield), as yellow to brown oil. For 19: ¹HNMR (300MHz, CDCl₃) δ 10.4 (s, 1H), 8.91 (s, 1H), 5.22 (m, 1H), 3.10 (m, 1H),2.87 (m, 1H), 2.48 (m, 1H), 2.09 (m, 1H), 0.92 (s, 9H), 0.21 (s, 3H),0.16 (s, 3H) ppm; LCMS (EI) m/z 312/314 (C₁₅H₂₂ClNO₂Si, (M+H)⁺).

Step 10.(R)-7-(tert-Butyldimethylsilyloxy)-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carbonitrile(20)

To a stirred solution of(R)-7-(tert-butyldimethylsilyloxy)-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carbaldehyde(19, 91.3 g, 292 mmol) in tetrahydrofuran (THF, 700 mL) at roomtemperature was added a solution of 14.8 M ammonia in water (350 mL,5950 mmol) and iodine (I₂, 80.0 g, 315 mmol) under an ice-water bath(the internal temperature was controlled at 16° C. to 22° C.). Theresulting reaction mixture was stirred at room temperature for 3 hoursbefore being quenched with a 10% Na₂S₂O₃ aqueous solution (200 mL). Themixture was extracted with ethyl acetate (2 500 mL). The combinedorganic extracts were washed with water (100 mL), dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure to give thedesired product,(R)-7-(tert-butyldimethylsilyloxy)-4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carbonitrile(20, 89.9 g, 98% yield) as yellow to brown oil. For 20: ¹H NMR (300 MHz,CDCl₃) δ 8.68 (s, 1H), 5.22 (m, 1H), 3.09 (m, 1H), 2.84 (m, 1H), 2.49(m, 1H), 2.07 (m, 1H), 0.92 (s, 9H), 0.20 (s, 3H), 0.18 (s, 3H) ppm;LCMS (EI) m/z 309/311 (C₁₅H₂₁ClN₂OSi, (M+H)⁺).

Example 6 Stability Study of Compound 1 Phosphoric Acid Salt Form I

Compound 1 phosphoric acid salt Form I was subjected to variousdifferent environmental conditions to assess stability. Results areshown in Table 2 below. As can be seen from the data, Form I has highstability even in the presence of heat and moisture. Only the heating at210° C. showed a possible conversion to another solid form, but theconversion was reversible upon exposure to 40° C./75% RH for 4 days.

TABLE 2 Crystalline Form Conditions (XRPD) Suspension in 0.5% Form Imethyl cellulose in water 40° C./75% RH for 1 week Form I 50° C. vacuumdry for Form I 1 day 100° C. vacuum dry Form I for 1 day 210° C. vacuumdry Not Form I for 20 min 210° C. vacuum Form I dry followed by 40°C./75% RH for 4 weeks

Example 8 Preparation of Compound 1 Phosphoric Acid Salt Form IProcedure A-Isopropanol:

To a solution of Compound 1 (25.68 mg, 0.05 mmol) in isopropanol (0.5mL) was added 0.056 mL of a 0.1 M solution of phosphoric acid (0.12mmol, 1.12 eq.) in isopropanol. The reaction mixture was stirredovernight. The resultant precipitate was collected by filtration, andthe filter cake was air-dried to yield Form I.

Procedure B-Acetonitrile:

Compound 1 (50.35 mg, 0.216 mmol, 1 eq.) was combined with 0.1 mL ofacetonitrile, and the mixture was stirred for 2 min to give a clearsolution. To the resultant solution was added 0.108 mL of 1M solution ofphosphoric acid (0.108 mmol, 1.1 eq.) in isopropanol to give a stickyslurry. The reaction mixture was heated to 78° C. and stirred for 2 h(note: slurry), after which time the reaction mixture was cooled to roomtemperature and stirred overnight. The resultant precipitate wascollected by filtration, and the filter cake was dried to give Form I.

The Karl Fischer titration indicated that the salt of Compound 1 fromacetonitrile contained about 1.419% water.

Procedure C-Ethanol:

Compound 1 (50.30 mg, 0.216 mmol, 1 eq.) was combined with 1.2 mL ofethanol, and the mixture was stirred to give a clear solution. To theresultant solution was added 0.108 mL of IM solution of phosphoric acid(0.108 mmol, 1.1 eq.) in isopropanol to give a slurry. The resultantreaction mixture was heated to 79° C. and stirred to 2 h, after whichtime the reaction mixture was stirred at 81-83° C. for 2 h (note:slurry). The reaction mixture was cooled to room temperature and stirredfor 2 h. The resultant precipitate was separated by filtration and thefilter cake was dried to give Form I (55 mg, 91.8%).

Procedure D-Methanol:

Compound 1 (50 mg, 0.216 mmol, 1 eq.) was combined with 0.5 mL ofmethanol to give a clear solution. To the resultant solution was added0.95 mL of 1M solution of phosphoric acid (0.95 mmol, 1.25 eq.) inisopropanol to give a sticky slurry. Methanol (0.5 mL) was added and themixture was stirred for 1 h, after which time the reaction mixture washeated to 78° C. and stirred at that temperature for 2 h (note: slurry).The resultant precipitate was collected by filtration and the filtercake was dried to give Form I (42.6 mg, 91.7%).

The stoichiometric ratio between Compound 1 free base and phosphoricacid was determined by elemental analysis as 1:1. Elemental analysis:Calculated for C₂₆H₂₉F₃N₅O₇P: C, 51.07; H, 4.78; N, 11.45; P, 5.07.Found: C, 49.23; H, 4.57; N, 10.85; P, 5.28.

Example 9 Stability Study of Compound 1 Phosphoric Acid Salt Form IUnder Phase Equilibration Conditions at 25° C. and 50° C.

Phase equilibration studies were designed to provide information on apredominant crystal form for phase identification. Compound 1 phosphoricacid salt (Form I) was equilibrated in a representative group ofsolvents at 25±1° C. (chloroform, DMF, 1,4-dioxane, methanol andmethanol/20% water, 2-methoxyethanol and 2-methoxyethanol/20% water,MIBK, THF and THF/20% water, acetone, n-BuOH and n-BuOH/20% water, EtOHand EtOH/20% water, isobutyl acetate, 1-propanol and 1-propanol/20%water, isopropanol, water, and MEK) and 50±1° C. (chloroform, DMF,1,4-dioxane, methanol and methanol/20% water, 2-methoxyethanol and2-methoxyethanol/20% water, MIBK, THF and THF/20% water, acetone, n-BuOHand n-BuOH/20% water, EtOH and EtOH/20% water, EtOAc, ethyl formate,1-propanol and 1-propanol/20% water, isopropanol, IPA/MeOH/Water(1.73/0.79/0.08), IPA/water (3/2), water) controlled by IKA® ETS-D5temperature controller and IKA® RCT basic safety control.

To 3 mL of a solvent or a mixture of solvents (chosen from a list forthe respective temperature) was added Compound 1 phosphate (Form I)until a cloudy solution was obtained, then, about 30 mg of Compound 1phosphate (Form I) was added to the cloudy solution. The mixture wasstirred at 25±1° C. or 50±1° C., respectively, for 2-3 days. The solidwas filtered and analyzed by XRPD. The material was Form I for phaseequilibration at 25±1° C. and 50±1° C. in all of the solvents andsolvent mixtures tested, which is the same as the starting materialCompound 1 phosphate (Form I).

Example 10 Preparation and Characterization of Compound 1 PhosphoricAcid Salt Form II.

A saturated solution of Compound 1 phosphate Form I (prepared in Example1, 20 mL) in DMF was evaporated under air without stirring at 25±1° C.to give a solid, which was characterized by XRPD, DSC and TGA as FormII.

Experimental parameters for acquiring the XRPD data are as described inExample 2. The XRPD pattern is shown in FIG. 7 and XRPD data is providedin Table 3.

TABLE 3 XRPD Data Form II 2-Theta (°) Height H % 4.7 2194 100 9.4 97044.2 13.1 82 3.7 14.1 188 8.6 16.2 83 3.8 18.8 2026 92.4 19.2 171 7.821.2 446 20.3 22.3 107 4.9 23.0 123 5.6 24.8 305 13.9 26.4 99 4.5 26.7139 6.3 27.6 113 5.1 28.4 83 3.8 29.0 29 1.3 29.4 40 1.8 30.2 184 8.433.3 322 14.7 34.1 135 6.2 34.9 264 12 38.2 136 6.2 38.8 48 2.2 39.8 853.9 43.3 166 7.6

Experimental parameters for acquiring the DSC data are as described inExample 3. The DSC thermogram is shown in FIG. 8 and displays anendothermic event at about 249° C.

Experimental parameters for acquiring the TGA data are as described inExample 4. The TGA thermogram is shown in FIG. 9.

Example 11 Preparation and Characterization of Compound 1 PhosphoricAcid Salt Form III.

A 20 mL saturated solution of Compound 1 phosphate (Form I) in DMF wasevaporated under air without string at 50±1° C. to give a solid whichwas characterized by XRPD, DSC and TGA as Form III.

Experimental parameters for acquiring the XRPD data are as described inExample 2. The XRPD pattern is shown in FIG. 10 and XRPD data isprovided in Table 4.

TABLE 4 XRPD Data Form III 2-Theta (°) Height H % 4.6 425 85.1 7.1 509.9 9.4 129 25.9 13.3 166 33.3 15.7 89 17.8 16.3 173 34.7 18.9 500 10019.2 289 57.9 21.2 290 58 22.5 220 44.1 23.1 216 43.2 24.3 88 17.7 24.9142 28.4 25.6 65 13.1 26.7 165 33.1 27.7 89 17.7 29.1 61 12.1 30.4 6212.4 33.4 41 8.2 34.2 66 13.2 35.0 72 14.3 38.3 46 9.3 38.8 57 11.5 43.443 8.7

Experimental parameters for acquiring the DSC data are as described inExample 3. The DSC thermogram is shown in FIG. 11 and displays anendothermic event at about 250° C.

Experimental parameters for acquiring the TGA data are as described inExample 4. The TGA thermogram is shown in FIG. 12.

Example 12 Preparation and Characterization of Compound 1 PhosphoricAcid Salt Form IV.

A 20 mL saturated solution of Compound 1 phosphate Form I in water wasevaporated under air without string at 50±1° C. to give a solid whichwas characterized by XRPD, DSC and TGA as Form IV.

Experimental parameters for acquiring the XRPD data are as described inExample 2. The XRPD pattern is shown in FIG. 13 and XRPD data isprovided in Table 5.

TABLE 5 XRPD Data Form IV 2-Theta (°) Height H % 4.1 81 62.1 4.9 52 39.96.9 45 34.7 7.4 63 48.3 8.0 37 28.6 11.0 44 34 12.8 38 29 13.3 96 73.916.4 130 100 17.7 78 59.4 18.1 88 67.1 18.6 102 77.8 19.0 55 42 19.8 11890.1 20.6 42 32.3 21.4 114 87 22.6 46 35.5 23.3 81 62 25.0 61 46.8 26.746 35.5 35.7 26 20.2 38.4 27 20.3

Experimental parameters for acquiring the DSC data are as described inExample 3. The DSC thermogram is shown in FIG. 14 and displays anendothermic event at about 245° C.

Experimental parameters for acquiring the TGA data are as described inExample 4. The TGA thermogram is shown in FIG. 15.

Example 13 Preparation and Characterization of Compound 1 PhosphoricAcid Salt Form V.

A 100 mL saturated solution of Compound 1 phosphate in water, preparedat 35° C., was quench-cooled to 4-5° C., and kept at the temperature for1 h to give a thin slurry which was filtered and air-dried for 1 h. Thesolid was assigned as Compound 1 phosphate Form V and characterized byXRPD, DSC and TGA.

Experimental parameters for acquiring the XRPD data are as described inExample 2. The XRPD pattern is shown in FIG. 16 and XRPD data isprovided in Table 6.

TABLE 6 XRPD Data Form V 2-Theta (°) Height H % 4.2 50 8.5 5.5 53 9 7.3301 51.4 9.3 75 12.8 10.2 46 7.8 10.9 316 53.9 11.9 50 8.5 12.7 72 12.413.1 148 25.2 14.7 84 14.4 16.4 479 81.8 17.5 159 27.2 18.1 232 39.618.5 586 100 19.8 531 90.6 20.6 88 15 21.2 102 17.3 22.6 244 41.6 23.191 15.5 23.8 81 13.9 24.7 120 20.6 26.1 252 43.1 26.7 161 27.5 30.5 11619.7 30.8 61 10.5 31.8 69 11.8 35.1 86 14.6 35.5 65 11.2 37.3 104 17.737.7 65 11.1 39.7 53 9.1 44.2 51 8.7

Experimental parameters for acquiring the DSC data are as described inExample 3. The DSC thermogram is shown in FIG. 17.

Experimental parameters for acquiring the TGA data are as described inExample 4. The TGA thermogram is shown in FIG. 18.

Example 14 Preparation and Characterization of Compound 1 PhosphoricAcid Salt Form VI.

To 150 mL water was added Compound 1 phosphate to give a slurry whichwas stirred for 2 h to give a suspension. The suspension was filteredand the filtrate was cooled to 4-5° C. and held at 4-5° C. for 3 days.The suspension was filtered to isolate crystal solid which wascharacterized by XRPD, DSC and TGA as Form VI.

Experimental parameters for acquiring the XRPD data are as described inExample 2. The XRPD pattern is shown in FIG. 19 and XRPD data isprovided in Table 7.

TABLE 7 XRPD Data Form VI 2-Theta (°) Height H % 3.7 133 21.2 4.1 22235.4 6.5 629 100 8.3 437 69.5 10.7 216 34.4 13.2 230 36.6 14.6 171 27.316.1 61 9.7 17.3 263 41.8 18.3 59 9.4 19.1 444 70.7 20.1 171 27.2 20.877 12.2 21.5 130 20.7 21.8 74 11.7 22.8 59 9.4 24.1 85 13.5 25.1 11418.1 26.6 108 17.2 27.1 74 11.7 27.8 83 13.2 30.8 93 14.8 34.5 47 7.536.2 80 12.7 38.5 60 9.5 40.2 54 8.7 40.8 49 7.8 44.6 45 7.2

Experimental parameters for acquiring the DSC data are as described inExample 3. The DSC thermogram is shown in FIG. 20.

Experimental parameters for acquiring the TGA data are as described inExample 4. The TGA thermogram is shown in FIG. 21.

Example 15

Study of Polymorphism of Compound 1 Phosphate using Anti-SolventAddition Saturated solutions of Compound 1 phosphate were prepared byadding the Compound 1 phosphate (Form I) to DMF, MeOH, MeOH/20% waterand DMSO respectively. An anti-solvent was added to induceprecipitation. MTBE, IPAc, EtOAc, MeCN, MIBK, MEK and toluene wereselected as the anti-solvents. Experiments that did not produce anyparticulate solids on anti-solvent addition were not studied further.

In antisolvent addition (see Table 8), Form I was identified fromMeOH/IPAc, MeOH/EtOAc, MeOH/MEK, aq. MeOH/MTBE, DMSO/MeCN, DMSO/IPAc,and DMSO/MEK. Form II was identified from MeOH/MeCN, aq. MeOH/MeCN, andaq. MeOH/IPAc. Amorphous was found from MeOH/TBME and MeOH/MIBK.

The XRPD pattern for amorphous Compound 1 phosphate is shown in FIG. 23.

TABLE 8 Solid Anti-solvent state Solvent (mL) (mL) form DMF (2 mL) IPAc(8 mL) n/a DMF (2 mL) MTBE (8 mL) n/a MeOH (2 mL) MTBE (8 mL) amorphousMeOH (2 mL) IPAc (8 mL) Form I MeOH (2 mL) EtOAc (8 mL) Form I MeOH (2mL) CH₂Cl₂ (8 mL) Clear solution MeOH (2 mL) MeCN (8 mL) Form II MeOH (2mL) MIBK (8 mL) amorphous MeOH (2 mL) MEK (8 mL) Form I MeOH/ (2 mL)MeCN (8 mL) Form II 20% water MeOH/ (2 mL) MTBE (8 mL) Form I 20% waterMeOH/ (2 mL) IPAc (8 mL) Form II 20% water DMSO (0.5 mL) MeCN (5 mL)Form I DMSO (0.5 mL) IPAc (5 mL) Form I DMSO (0.5 mL) MEK (5.5 mL) FormI DMSO (0.5 mL) Acetone (8 mL) n/a DMSO (0.5 mL) Toluene (6 mL) n/a

Example 16 Study of Polymorphism of Compound 1 Phosphate Using ReverseAddition

Saturated solutions of Compound 1 phosphate were prepared in DMF, MeOH,MeOH/20% water and DMSO listed in Table 9, and added to a larger volumeof a miscible anti-solvent (i.e. MTBE, IPAc, EtOAc, MeCN). Experimentsthat did not produce any particulate solids upon addition to theanti-solvent were not studied further. Form I was identified from mostsolvents, except Form II was identified from MeOH/MeCN, aq. MeOH/MeCN,and aq. MeOH/IPAc. Amorphous solid was identified from MeOH/MTBE,MeOH/EtOAc and MeOH/MIBK.

TABLE 9 Solid Anti-solvent state Solvent (mL) (mL) form DMF (2 mL) IPAc(8 mL) n/a DMF (2 mL) MTBE (8 mL) n/a MeOH (2 mL) MTBE (8 mL) AmorphousMeOH (2 mL) IPAc (8 mL) Form I MeOH (2 mL) EtOAc (8 mL) amorphous MeOH(2 mL) CH₂Cl₂ (8 mL) n/a MeOH (2 mL) MeCN (8 mL) Form II MeOH (2 mL)MIBK (8 mL) Amorphous MeOH (2 mL) MEK (8 mL) Form I MeOH/ (2 mL) MeCN (8mL) Form II 20% water MeOH/ (2 mL) MTBE (8 mL) Form I 20% water MeOH/ (2mL) IPAc (8 mL) Form II 20% water DMSO (0.5 mL) IPAc (5 mL) Form I DMSO(0.5 mL) MeCN (5 mL) Form I DMSO (0.5 mL) MEK (5.5 mL) Form I DMSO (0.5mL) Acetone (8 mL) n/a DMSO (0.5 mL) Toluene (6 mL) n/a

Example 17 Compound 1 Phosphate Study Using Quench-Cooling

Saturated solutions of Compound 1 phosphate prepared at 35° C. werequench cooled to about −20° C. to −25° C. to induce precipitation ofhigher energy crystalline forms. Representative solvents were chosenbased on solubility data measured at 25° C. and 50° C. (See Example 7).The studied solvents and the crystalline forms resulting from theexperiments are shown in Table 10. Form I was identified from aq. THFand aq. EtOH. Form V and Form VI were identified from water.

TABLE 10 Solvent (mL) Solid state form DMF Clear solution MeOH Clearsolution MeOH/20% water Clear solution 2-methoxyethanol Clear solution2-methoxyethanol/20% water Clear solution THF/20% water Form I EtOH/20%water Form I 1-PrOH/20% water Clear solution 2-PrOH/20% water Clearsolution Water (Cooled to 4-5° C.) Form V Water (Cooled to 4-5° C.) FormVI

Example 18 Study of Polymorphism of Compound 1 Phosphate Using Heatingand Cooling Cycles

This experiment was designed to search for stable crystalline forms.Saturated solutions were prepared at 50° C., and cooled in a bath slowlyby using a programmed circulating bath to give clear solution for allsolvents. To the clear solution was added about 10 mg of Compound 1phosphate Form I to give slurry. The resulting slurry was then heated to50° C. over 2 hours and then cooled down to 5° C. over 2 hours. Thisprocess was repeated for 3 days and the solid was filtered for furtheranalysis. The results are presented in Table 11. Form I was identifiedfor all of the samples.

TABLE 11 Solid Solvent (mL) state form DMF Form I Methanol Form IMethanol/20% water Form I 2-Methoxyethanol Form I 2-Methoxyethanol/20%water Form I THF/20% water Form I n-BuOH/10% water Form I EtOH/20% waterForm I n-Propanol/20% water Form I IPA/30% water Form I Water Form I

Example 19 Study of Polymorphism of Compound 1 Phosphate UsingEvaporation

Evaporation studies were carried out to identify the predominant crystalform during uncontrolled precipitation. Experiments that did not resultin any particulate solids (i.e. clear thin films and oils) were notstudied further (n/a). XRPD was used to characterize the solid-statemorphology of the crystalline forms of the evaporation samples at 25±1°C. and 50±1° C. controlled by IKA® ETS-D5 temperature controller andIKA® RCT basic safety control.

The results are presented in Table 12 (25±1° C.) and Table 13 (50±1°C.). Evaporation at 25±1° C. (Table 12) resulted in polymorphic Form II(DMF). Evaporation at 50±1° C. (Table 13) resulted in two polymorphicforms including Form III (DMF) and Form IV (water).

TABLE 12 Evaporation at 25 ± 1° C. Solvent (mL) Solid state form MeCNn/a Chloroform n/a Dichloromethane n/a DMF Form II 1,4-Dioxane/5% watern/a 1,4-Dioxane n/a Methanol n/a Methanol/20% water Form I2-Methoxyethanol Form I MIBK n/a Toluene n/a Hexane n/a THF n/a Acetonen/a n-BuOH n/a MTBE n/a DMSO Form I EtOH n/a EtOAc n/a Ethyl formate n/aHeptane n/a Isobutyl acetate n/a IPAc n/a 1-Propanol n/a IPA n/a Watern/a MEK n/a

TABLE 13 Evaporation 50 ± 1° C. Solvent (mL) Solid state form MeCN n/aChloroform n/a DMF Form III 1,4-Dioxane n/a Methanol Form I MeOH/20%water Form I 2-Methoxyethanol Form I MIBK n/a Toluene n/a Hexane n/a THFn/a Acetone n/a n-BuOH n/a MTBE n/a DMSO Form I EtOH n/a EtOAc n/a Ethylformate n/a Heptane n/a Isobutyl acetate n/a IPAc n/a 1-Propanol n/a IPAn/a Water Form IV MEK n/a

Example 20 Competitive Stability Study of Compound 1 PhosphateCrystalline Solids in IPA/MeOH/Water/DMSO

To evaluate the transformation of Compound 1 phosphate solid forms,competitive slurry experiments were performed as follows: to a saturatedsolution (1.5 mL) of Compound 1 phosphate Form I in the solvent aslisted in Table 14 was added Form I (6 mg), followed by stirring to givea cloudy solution, then 6 mg each of Form II through Form VI were addedto the mixture. The slurry was stirred for 2 days at room temperatureand analyzed by XRPD. The results in Table 14 revealed that the Compound1 phosphate Form I appears to be the most stable form in either of theIPA/methanol/water/DMSO mixtures.

TABLE 14 Solvent (mL) Solid state form IPA/MeOH/water/DMSO(61.8/28.2/2.9/7.1) Form I IPA/MeOH/water/DMSO (56.6/28.3/3.0/12.1) FormI

Example 22 Karl Fisher Titration of the Compound 1 Phosphate CrystallineSolid Forms

The results of Karl Fisher titration of the Compound 1 phosphatepolymorphs Forms I-VI are presented in Table 15.

TABLE 15 Solid state form Water % Notes Form I 1.5 GMP dried Form II 1.4Sample sealed in vial Form III 1.4 Sample sealed in vial Form IV 3.0Sample sealed in vial Form V 2.4 Sample sealed in vial Form VI 15.9Almost fresh sample

Karl-Fischer analysis of Compound 1 phosphate Forms I-VI was conducted.Each experiment revealed a water content within the range of 1.40-15.9%indicating that each of the Forms I-VI may be hydrated.

Example 23 Preparation and Characterization of Compound 1 DihydrochloricAcid Salt

Compound 1 (55.2 mg, 0.107 mmol) was combined with 0.7 mL isopropylalcohol (IPA) and stirred for 2 minutes to give a clear solution.Hydrochloric acid solution (0.25 mL, 0.25 mmol, 2.34 eq; 1M HCl inIPA/IPAc from 3.7 M HCl in IPAc) was added to give a slurry which washeated to 50° C. and stirred for 15 min. The resulting mixture wascooled to RT, stirred overnight, filtered and dried under vacuum (6 h)to give the final product (61.8 mg, 98%).

The stoichiometric ratio between Compound 1 and hydrochloric acid wasdetermined by elemental analysis as 1:2. ¹H NMR indicated the saltcontained 7.8% isopropanol, and Karl-Fischer titration indicated a watercontent of about 0.5⁸⁶%. Elemental Analysis: Calc'd for C₂₉H₃₅Cl₂F₃N₅O₄:C, 53.67; H, 5.35; N, 10.80; Cl, 10.95. Found: C, 53.26; H, 5.21; N,10.57; Cl, 10.83.

Compound 1 dihydrochloric acid salt was characterized by XRPD, DSC andTGA. Experimental parameters for acquiring the XRPD data are asdescribed in Example 2. The XRPD pattern is shown in FIG. 4.Experimental parameters for acquiring the DSC data are as described inExample 3. The DSC thermogram is shown in FIG. 5. The DSC thermogramrevealed a major endothermic peak at about 213° C. Experimentalparameters for acquiring the TGA data are as described in Example 4. TheTGA thermogram is shown in FIG. 6.

XRPD data is provided in Table 17.

TABLE 17 2-Theta (°) Height H % 3.8 176 19.5 8.3 376 41.8 11.2 152 16.912.4 231 25.7 13.9 79 8.8 15.6 218 24.2 16.6 129 14.3 18.9 632 70.3 20.0103 11.4 22.1 186 20.6 23.1 194 21.5 23.9 108 12.1 25.0 899 100 26.2 9510.6 27.2 167 18.6 28.4 197 21.9 30.0 228 25.3 31.8 150 16.7 33.6 16318.2 35.2 184 20.5 37.3 150 16.7 39.6 69 7.6 41.9 148 16.5 43.3 63 7

Example 24 Preparation and Characterization of Compound 1Monohydrochloric Acid Salt

To a solution of Compound 1 free base (see Example 1, step 6; 0.05 mmol,25.68 mg) in isopropanol (0.5 mL, 0.1 M) was added 0.056 mL ofhydrochloric acid (0.056 mmol, 1.12 eq., 1.0 M solution in IPA/IPAcprepared from 3.7 M HCl in IPAc (isopropyl acetate)). The reactionmixture was stirred overnight. The resultant precipitate was removed byfiltration, and the filter cake was washed with MTBE and the solid wasdried under vacuum overnight to afford the title salt.

Compound 1 monohydrochloric acid salt was characterized by XRPD and DSC.An XRPD pattern is provided in FIG. 22. Experimental parameters foracquiring the DSC data are as described in Example 3. The DSC thermogramis shown in FIG. 23.

XRPD data is provided in Table 18.

TABLE 18 2-Theta (°) Height H % 3.5 102 27.3 4.0 105 28 7.8 126 33.6 8.8134 35.7 11.9 57 15.3 12.6 121 32.2 14.5 138 36.8 15.0 47 12.5 15.9 7018.8 16.6 44 11.6 17.4 113 30.2 19.7 74 19.8 20.4 59 15.8 20.6 54 14.422.3 74 19.9 23.8 186 49.6 24.6 84 22.3 25.2 375 100 26.3 80 21.4 26.582 21.8 27.4 38 10.2 28.8 90 24 29.2 72 19.2 31.1 75 19.9 31.5 44 11.731.7 42 11.3 34.5 69 18.3 37.6 46 12.4 40.9 43 11.4 41.3 32 8.6 41.8 348.9

Example 25 Preparation and Characterization of Compound 1 Maleic AcidSalt

A volume of 1.0 mL of isopropanol was added to Compound 1 free base(50.30 mg, 0.216 mmol, 1 eq.). The resultant mixture was stirred to givea clear solution. Maleic acid (14.2 mg. 0.122 mmol, 1.21 eq.) was addedto this solution and the resultant reaction mixture was stirred to givea clear solution. The stirring continued for 1 h. To this solution, 1 mgof crystals (seeds) obtained from IPA/heptane were added and theresultant mixture was stirred to give a slurry. The slurry wascontinuously stirred for 3 h. The precipitate was removed by filtration,and the filter cake was washed with MTBE and dried under vacuumovernight to afford the title salt (56.8 mg, 89.2%).

The stoichiometric ratio of the salt between Compound 1 free base andmaleic acid was determined by ¹H NMR as 1:1. The crystallinity of theCompound 1 maleate was confirmed by XRPD, DSC, and TGA.

Experimental parameters for acquiring the XRPD data are as described inExample 2. The XRPD pattern is shown in FIG. 24. Experimental parametersfor acquiring the DSC data are as described in Example 3. The DSCthermogram is shown in FIG. 25. Experimental parameters for acquiringthe TGA data are as described in Example 4. The TGA thermogram is shownin FIG. 26.

XRPD data is provided in Table 19.

TABLE 19 2-Theta (°) Height H % 5.2 108 3.9 9.0 762 27.4 9.5 851 30.610.4 504 18.1 11.2 1255 45.1 12.8 204 7.3 14.8 632 22.7 15.9 748 26.917.0 323 11.6 17.8 460 16.5 18.5 2781 100 19.5 687 24.7 19.9 1036 37.220.9 1007 36.2 21.3 1421 51.1 22.9 2122 76.3 23.6 538 19.3 24.4 575 20.724.8 1904 68.5 25.8 1638 58.9 27.6 1166 41.9 29.2 492 17.7 30.9 102536.9 32.5 268 9.7 33.4 97 3.5 34.2 429 15.4 35.6 252 9 36.0 152 5.5 36.7200 7.2 37.6 115 4.2 38.2 61 2.2 38.6 77 2.8 39.9 123 4.4 40.7 220 7.941.6 251 9 42.3 471 16.9 43.2 328 11.8 43.6 161 5.8

Example 26 Preparation and Characterization of Compound 1 Adipic AcidSalt

A volume of 0.6 mL of isopropanol was added to Compound 1 free base(37.8 mg, 0.216 mmol. 1 eq) and the resultant mixture was stirred for 1min to give a clear solution. Adipic acid (26.8 mg, 0.183 mmol, 2.49eq.) was added to the solution and the resultant slurry was stirred atroom temperature for 5 min. The reaction mixture was heated to 50° C.and stirred at that temperature for 15 min. (Note: clear solution). Thereaction mixture was cooled to room temperature and stirred for 3 h.Heptane (0.2 mL) was added and the reaction mixture was stirred to givea slurry, which was continuously stirred overnight. The precipitate wasremoved by filtration and the filter cake was washed with MTBE,collected, and dried under vacuum overnight to afford the title salt(36.5 mg, 84.5% yield).

The stoichiometric ratio of the salt between Compound 1 free base andadipic acid was determined by ¹H NMR as 1:1. The crystallinity ofCompound 1 adipate was confirmed by XRPD, DSC, and TGA. Experimentalparameters for acquiring the XRPD data are as described in Example 2.The XRPD pattern is shown in FIG. 27. Experimental parameters foracquiring the DSC data are as described in Example 3. The DSC thermogramis shown in FIG. 28. Experimental parameters for acquiring the TGA dataare as described in Example 4. The TGA thermogram is shown in FIG. 29.

XRPD data is provided in Table 20.

TABLE 20 2-Theta (°) Height H % 3.8 169 10 9.0 443 26.3 9.3 1025 60.810.1 235 14 12.0 267 15.8 12.5 62 3.7 13.3 110 6.5 15.0 754 44.7 16.2464 27.5 17.6 368 21.8 18.7 792 46.9 20.0 1687 100 21.2 147 8.7 22.1 70842 22.7 415 24.6 24.3 1154 68.4 24.9 766 45.4 26.9 254 15.1 27.1 42525.2 28.7 337 20 30.2 75 4.5 31.7 124 7.3 33.7 140 8.3 35.0 95 5.6 35.658 3.4 36.3 71 4.2 36.7 77 4.6 38.2 119 7 40.5 62 3.7 41.8 168 9.9 43.2100 5.9

Example 27 Preparation and Characterization of Compound 1 HydrobromicAcid Salt

To a 0.1 M solution of Compound 1 free base (0.05 mmol, 25.68 mg) inisopropanol (0.5 mL) was added 0.12 mL of hydrobromic acid (0.12 mmol,2.4 eq., 1.0 M solution in isopropanol/water). The resultant mixture wasstirred overnight to give a slurry. The precipitate was removed byfiltration and the filter cake was washed with MTBE and dried undervacuum overnight to give the desired product.

Experimental parameters for acquiring the XRPD data are as described inExample 2. The XRPD pattern is shown in FIG. 30. Experimental parametersfor acquiring the DSC data are as described in Example 3. The DSCthermogram is shown in FIG. 31. Experimental parameters for acquiringthe TGA data are as described in Example 4. The TGA thermogram is shownin FIG. 32.

XRPD data is provided in Table 21.

TABLE 21 2-Theta (°) Height H % 3.8 171 55.4 4.2 142 46.2 6.5 99 32.39.5 93 30.3 10.1 41 13.2 10.9 47 15.1 12.9 189 61.3 13.2 102 33.2 13.964 20.6 15.6 69 22.4 16.6 263 85.3 17.9 143 46.6 18.9 48 15.6 19.5 20165.2 19.9 77 24.9 21.7 101 33 22.5 308 100 23.0 111 35.9 23.7 179 58.224.3 188 61 26.5 256 83.1 27.5 239 77.5 28.3 135 43.8 30.6 48 15.5 32.549 15.9 33.6 77 25.1 33.9 86 27.8 35.3 50 16.3

Example 28 Preparation and Characterization of Compound 1 Mandelic AcidSalt

To the 0.1 M solution of Compound 1 free base (0.05 mmol, 25.68 mg) inIPA (0.5 mL) was added mandelic acid (8.1 mg, 0.053 mmol, 1.06 eq.). Themixture was stirred overnight. The slurry was filtered, washed with MTBEto give Compound 1 mandelate salt, which was analyzed by XRPD, DSC andTGA.

Experimental parameters for acquiring the XRPD data are as described inExample 2. The XRPD pattern is shown in FIG. 33. Experimental parametersfor acquiring the DSC data are as described in Example 3. The DSCthermogram is shown in FIG. 34. Experimental parameters for acquiringthe TGA data are as described in Example 4. The TGA thermogram is shownin FIG. 35.

XRPD data is provided in Table 22.

TABLE 22 2-Theta (°) Height H % 4.0 190 9.7 4.8 54 2.7 8.7 89 4.5 11.2753 38.3 12.0 286 14.5 13.8 207 10.6 14.9 359 18.3 15.7 142 7.2 16.7 20610.5 17.3 222 11.3 18.6 788 40.1 20.2 107 5.4 20.6 379 19.3 22.5 53727.3 24.1 1965 100 25.3 338 17.2 26.6 221 11.3 27.9 118 6 28.6 162 8.229.4 124 6.3 30.2 60 3 31.0 138 7 31.5 59 3 32.7 91 4.6 33.5 110 5.635.7 125 6.4 37.5 103 5.2 38.6 129 6.6 39.3 126 6.4 40.9 112 5.7 42.3 854.4 44.0 113 5.7

Example 29 Compound 1 Salicylic Acid Salt

To the 0.1 M solution of Compound 1 free base (0.05 mmol, 25.68 mg) inisopropyl alcohol (IPA) (0.5 mL) was added salicylic acid (0.058 mmol,8.01 mg, 1.16 eq.). The resultant reaction mixture was stirred overnightto form a slurry. The precipitate was removed by filtration, and thefilter cake was washed with methyl tert-butyl ether (MTBE) to giveCompound 1 salicylic acid salt, which was characterized by XRPD and DSC.

Experimental parameters for acquiring the XRPD data are as described inExample 2. The XRPD pattern is shown in FIG. 36. Experimental parametersfor acquiring the DSC data are as described in Example 3. The DSCthermogram is shown in FIG. 37.

XRPD data is provided in Table 23.

TABLE 23 2-Theta (°) Height H % 8.7 58 2.7 9.7 77 3.5 10.9 234 10.7 11.3406 18.7 11.8 609 28 13.2 80 3.7 13.6 48 2.2 14.5 163 7.5 14.8 366 16.815.6 231 10.6 16.7 288 13.2 18.3 143 6.6 18.7 422 19.4 19.9 161 7.4 20.658 2.7 21.2 735 33.8 21.9 1536 70.6 23.0 1123 51.6 23.4 2176 100 24.1716 32.9 25.1 501 23 25.8 93 4.3 26.3 410 18.8 27.1 57 2.6 27.7 151 6.928.1 142 6.5 29.4 243 11.1 29.9 63 2.9 31.3 131 6 32.9 110 5 35.7 84 3.936.5 120 5.5 36.8 231 10.6 37.9 170 7.8 39.4 57 2.6 42.0 185 8.5 42.6139 6.4 44.0 66 3 44.4 62 2.9

Example 30

Compound 1 Benzoic Acid Salt To the 0.1 M solution of Compound 1 freebase (0.05 mmol, 25.68 mg) in isopropyl alcohol (IPA) (0.5 mL) was addedbenzoic acid (7.05 mg, 0.0577 mmol, 1.16 eq.). The resultant reactionmixture was stirred overnight to form a slurry. The precipitate wasremoved by filtration, and the filter cake was washed with methyltert-butyl ether (MTBE) to give Compound 1 benzoic acid salt, which wasanalyzed by XRPD.

Experimental parameters for acquiring the XRPD data are as described inExample 2. The XRPD pattern is shown in FIG. 38.

XRPD data is provided in Table 24.

TABLE 24 2-Theta (°) Height H % 4.9 156 15.9 8.7 83 8.5 9.8 180 18.310.7 129 13.1 11.6 566 57.8 13.1 74 7.5 14.9 225 22.9 15.9 66 6.7 16.9389 39.7 18.8 346 35.4 21.5 692 70.7 23.2 862 88 23.7 980 100 24.9 56657.8 26.2 243 24.9 27.2 168 17.2 27.5 139 14.1 29.2 78 8 31.2 113 11.532.7 145 14.8 35.3 128 13.1 36.4 62 6.3 37.3 120 12.3 38.3 85 8.7 39.9130 13.3 42.5 187 19

Example 31 Compound 1 Benzenesulfonic Acid Salt

To the 0.1 M solution of Compound 1 free base (0.05 mmol, 25.68 mg) inisopropyl alcohol (IPA) (0.5 mL) was added benzenesulfonic acid (0.11 mLof 0.5 M in isopropyl alcohol, 0.055 mmol, 1.1 eq.). The resultantreaction mixture was stirred overnight to give a slurry. The precipitatewas removed by filtration to give Compound 1 besylate salt, which wascharacterized by XRPD.

Experimental parameters for acquiring the XRPD data are as described inExample 2. The XRPD pattern is shown in FIG. 39.

XRPD data is provided in Table 25.

TABLE 25 2-Theta (°) Height H % 3.9 156 20 6.6 406 52.1 9.1 188 24.112.0 109 14 12.9 306 39.2 13.3 267 34.3 14.5 237 30.4 15.7 56 7.1 16.2162 20.7 18.0 780 100 19.2 197 25.2 19.8 109 14 20.5 94 12 21.2 125 16.121.8 182 23.3 23.5 356 45.7 23.9 522 66.9 25.2 56 7.1 26.1 79 10.1 28.489 11.4 29.7 148 18.9 31.6 65 8.4 32.9 46 5.8 35.9 45 5.7 36.4 69 8.839.8 47 6 42.8 71 9.1 43.7 52 6.6

Example 32 Compound 1 L-Pyroglutamic Acid Salt

To the 0.1 M solution of Compound 1 free base (0.05 mmol, 25.68 mg) inisopropyl alcohol (IPA) (0.5 mL) was added L-pyroglutamic acid (7.25 mg,0.056 mmol, 1.12 eq.). The resultant reaction mixture was stirred for 5h, after which time heptane (0.3 mL) was added. The reaction mixture wascontinuously stirred overnight to form a slurry. The precipitate wasremoved by filtration, and the filter cake was washed with methyltert-butyl ether (MTBE) to give Compound 1 L-pyroglutamic acid salt,which was characterized by XRPD.

Experimental parameters for acquiring the XRPD data are as described inExample 2. The XRPD pattern is shown in FIG. 40.

XRPD data is provided in Table 26.

TABLE 26 2-Theta (°) Height H % 4.4 344 32.7 8.3 49 4.6 9.2 86 8.2 10.7343 32.6 11.5 1052 100 12.5 74 7.1 13.5 176 16.7 15.0 102 9.7 15.4 20219.2 16.1 100 9.5 17.5 73 6.9 18.0 365 34.7 19.0 145 13.8 19.8 72 6.820.7 576 54.7 21.2 706 67.1 22.9 539 51.2 23.7 66 6.3 24.5 139 13.2 25.298 9.3 26.4 124 11.8 29.2 158 15 32.4 85 8.1 33.0 47 4.4 34.0 49 4.641.3 51 4.9 41.9 46 4.4

Example 33 Compound 1 Methanesulfonic Acid Salt

To the 0.1 M solution of Compound 1 free base (0.05 mmol, 25.68 mg) inisopropyl alcohol (IPA) (0.5 mL) was added methanesulfonic acid (0.055mmol, 1.1 eq., 0.055 mL of 1.0 M solution in EtOH). The resultantreaction mixture was stirred for 5 h, after which time heptane (0.3 mL)was added to the reaction mixture. The reaction mixture was continuouslystirred for 24 h to give a slurry. The precipitate was removed byfiltration to give Compound 1 mesylate salt, which was analyzed by XRPD.

Experimental parameters for acquiring the XRPD data are as described inExample 2. The XRPD pattern is shown in FIG. 41.

XRPD data is provided in Table 27.

TABLE 27 2-Theta (°) Height H % 3.6 143 18.6 6.4 60 7.8 9.6 55 7.1 10.444 5.7 12.7 222 28.9 13.5 650 84.6 14.7 528 68.7 15.1 135 17.6 16.7 59877.8 18.6 425 55.3 19.3 697 90.7 20.0 697 90.8 20.7 434 56.5 21.6 8110.6 22.1 214 27.9 22.4 146 19 23.0 295 38.5 24.1 447 58.2 24.4 768 10024.8 234 30.4 25.7 354 46.1 26.8 434 56.5 27.2 715 93.1 28.1 364 47.329.4 240 31.3 30.1 236 30.7 31.5 138 17.9 32.1 140 18.2 34.1 236 30.835.0 94 12.2 35.4 71 9.2 36.2 120 15.6 37.4 196 25.5 38.0 73 9.5 38.8183 23.8 39.8 90 11.7 40.1 61 8 40.4 59 7.7 41.0 76 9.9 41.4 54 7 41.970 9.1 42.1 49 6.4 42.9 67 8.7 43.4 108 14

Example 34

Compound 1 (1S)-(+)-10-camphorsulfonic Acid Salt

To the 0.1 M solution of Compound 1 free base (0.05 mmol, 25.68 mg) inisopropyl alcohol (IPA) (0.5 mL) was added (1S)-(+)-10-camphorsulfonicacid (0.055 mmol, 1.1 eq., 0.11 mL of 0.5 M solution in IPA) (CASregistry number 3144-16-9; Aldrich catalog number C2107-500G). Theresultant reaction mixture was stirred for 5 h, after which time heptane(0.3 mL) was added to the reaction mixture. The reaction mixture wascontinuously stirred overnight to give a slurry. The precipitate wasremoved by filtration to give Compound 1 camsylate salt, which wasanalyzed by XRPD.

Experimental parameters for acquiring the XRPD data are as described inExample 2. The XRPD pattern is shown in FIG. 42.

XRPD data is provided in Table 28.

TABLE 28 2-Theta (°) Height H % 3.7 140 25.9 6.6 232 43 7.1 539 100 10.9249 46.1 13.6 249 46.2 16.1 292 54.1 17.7 372 69 18.8 188 34.8 19.9 50092.7 21.3 70 12.9 23.2 321 59.6 25.6 90 16.7 28.4 124 22.9 29.6 62 11.531.7 60 11.1 35.1 99 18.4 43.5 64 11.8

Example 35 Compound 1 Fumaric Acid Salt

To the 0.1 M solution of Compound 1 free base (0.05 mmol, 25.68 mg) inisopropyl alcohol (IPA) (0.5 mL) was added maleic acid (6.69 mg, 0.058mmol, 1.16 eq.). The reaction mixture was stirred for 5 h, after whichtime heptane (0.3 mL) was added to the reaction mixture. The reactionmixture was continuously stirred overnight to give a slurry. Theprecipitate was removed by filtration to give Compound 1 fumarate salt,which was analyzed by XRPD.

Compound 1 fumaric acid salt was characterized by XRPD. Experimentalparameters for acquiring the XRPD data are as described in Example 2.The XRPD pattern is shown in FIG. 43.

XRPD data is provided in Table 29.

TABLE 29 2-Theta (°) Height H % 3.8 397 77.6 5.2 120 23.4 7.3 511 10010.4 89 17.4 11.6 74 14.5 13.2 145 28.4 16.2 68 13.3 17.1 126 24.6 20.8239 46.7 24.3 216 42.2 25.6 81 15.9 27.8 78 15.3 31.1 71 13.9 41.9 5210.1

Example 36 Compound 1 Sulfuric Acid Salt

To the 0.1 M solution of Compound 1 free base (0.05 mmol, 25.68 mg) inIPA (0.5 mL) was added sulfuric acid (0.055 mmol, 1.1 eq., 0.055 mL of1.0 M solution in IPA). The reaction mixture was stirred overnight togive a slurry. The precipitate was removed by filtration to giveCompound 1 sulfate salt, which was characterized by XRPD.

Compound 1 sulfuric acid salt was characterized by XRPD. Experimentalparameters for acquiring the XRPD data are as described in Example 2.The XRPD pattern is shown in FIG. 44.

XRPD data is provided in Table 30.

TABLE 30 2-Theta (°) Height H % 3.6 185 100 6.3 169 91.8 8.7 89 48.310.8 54 29.1 12.8 51 27.6 13.7 57 31.1 15.4 132 71.7 19.0 152 82.1 20.2114 62 21.2 85 46 21.6 114 61.9 23.1 76 40.9 24.0 62 33.4 24.8 68 36.625.4 59 31.8 27.1 110 59.6 27.2 111 60.4

Example 37 Compound 1 L-Tartaric Acid Salt

To the 0.1 M solution of Compound 1 free base (0.05 mmol, 25.68 mg) inisopropyl alcohol (IPA) (0.5 mL) was added L-tartaric acid (8.71 mg,0.058 mmol, 1.16 eq.). The reaction mixture was stirred for 5 h, afterwhich time heptane (0.3 mL) was added to the reaction mixture. Thereaction mixture was continuously stirred overnight to give a slurry.The precipitate was removed by filtration to give Compound 1 L-tartratesalt, which was analyzed by XRPD.

Compound 1 L-tartaric acid salt was obtained as amorphous solid.Experimental parameters for acquiring the XRPD data are as described inExample 2. The XRPD pattern is shown in FIG. 45.

Example 38 Compound 1 D-Tartaric Acid Salt

To the 0.1 M solution of Compound 1 free base (0.05 mmol, 25.68 mg) inIPA (0.5 mL) was added D-tartaric acid (8.64 mg, 0.058 mmol, 1.16 eq.).The reaction mixture was stirred for 5 h, after which time heptane (0.3mL) was added to the reaction mixture. The reaction mixture wascontinuously stirred overnight to give a slurry. The precipitate wasremoved by filtration to give Compound 1 D-tartrate salt, which wasanalyzed by XRPD.

Compound 1 D-tartaric acid salt was obtained as amorphous solid (theXRPD pattern is shown in FIG. 46).

Example A Pim Enzyme Assays

Pim-1 and Pim-3 kinase assays-20 μL reactions can be run in white 384well polystyrene plates dotted with 0.8 μL compound/DMSO in the assaybuffer (50 mM Tris, pH 7.5, 0.01% Tween-20, 5 mM MgCl₂, 0.01% BSA, 5 mMDTT), containing 0.05 μM Biotin-labeled BAD peptide substrate (AnaSpec62269), 1 mM ATP, and 2.5 pM (Pim-1, Invitrogen PV3503) or 1.25 pM(Pim-3, Millipore 14-738) enzyme for 1 h at 25° C. Reactions are stoppedby addition of L STOP Buffer (150 mM Tris, pH=7.5, 150 mM NaCl, 75 mMEDTA, 0.01% Tween-20, 0.3% BSA) supplemented with Phospho-Bad (Ser112)Antibody (Cell Signaling 9291) diluted 666-fold, and Streptavidin donorbeads (PerkinElmer 6760002) along with Protein-A acceptor beads(PerkinElmer 6760137) at 15 μg/mL each. Supplementation of the STOPbuffer with beads and stopping the reactions are done under reducedlight. Prior to the stopping reactions STOP buffer with beads ispre-incubated for 1 h in the dark at room temperature. After stoppingthe reactions, plates are incubated for 1 h in the dark at roomtemperature before reading on a PHERAstar FS plate reader (BMG Labtech)under reduced light.

Pim-2 kinase assay-20 μL reactions are run in white 384 well polystyreneplates dotted with 0.8 μL compound/DMSO in the assay buffer (50 mM Tris,pH 7.5, 0.01% Tween-20, 5 mM MgCl₂, 0.01% BSA, 5 mM DTT), containing0.05 μM Fluorescein-labeled CREBtide peptide substrate (InvitrogenPV3508), 1 mM ATP, and 1 nM enzyme (Invitrogen PV3649) for 2 h at 25° C.Reactions are stopped by addition of 10 μL TR-FRET Dilution Buffer(Invitrogen PV3574) with 30 mM EDTA and 1.5 nM LanthaScreen Tb-CREBpSer133 antibody (Invitrogen PV3566). After 30 min. incubation at roomtemperature, plates are read on a PHERAstar FS plate reader (BMGLabtech).

Compounds or salts of the invention having an IC₅₀ of 2 μM or less whentested for PIM kinase activity under the assay conditions disclosedabove are considered active. Compound 1 was tested according to thisassay and found to have an IC₅₀<100 nM. Compound 1 phosphate salt andCompound 1 dihydrochloric acid salt were tested according to this assayand data is provided below in Table 16.

Although the above in vitro assays are conducted at 1 mM ATP, compoundscan also be evaluated for potency and in vitro activity against PIMtargets utilizing K_(m) conditions, where the concentration of ATP isset to the K_(m) value and the assay is more sensitive to PIM inhibitionactivity.

Example B Pim Cellular and Whole Blood Assays Pim Cell ProliferationAssays

KMS12BM and MOLM16 cells were purchased from DSMZ (Germany) and weremaintained according to the recommendations of suppliers. To measure theanti-proliferation activities of test compounds, the cells were platedin their respective culture medium (2×103 cells/200 μL/well) into96-well ultralow binding plates (Corning), with or without testcompound(s). After 3 to 4 days, [3H]-thymidine (1 μCi/well)(PerkinElmer) in PBS (10 μL) was then added to the cell culture for anadditional 12 hours before the incorporated radioactivity was separatedby filtration with water through GF/B filters (Packard Bioscience) andmeasured by liquid scintillation counting with a TopCount (PackardBioscience).

Pim pBAD Signaling Assays

To measure the effect of test compounds on the level of pBAD in cells,KMS12BM cells (DSMZ, Germany) were plated with RPMI and 10% FBS (4×10⁵cells/well/100 μL) into 96-well v-bottom polypropylene plates (Greiner)in the presence or absence of 5 μL of a concentration range testcompound(s). After 2.5 hours at 37° C. and 5% CO₂, the cells were lysedin 100 μL of cell extraction buffer (Cell Signaling Technology)containing PMSF, HALT, and protease inhibitors (Thermo, EMD Calbiochem).pBAD protein in the cell lysates was quantified with the Human pBAD S112ELISA Kit (Cell Signaling Technology). IC₅₀ determination was performedby fitting the curve of percent inhibition versus the log of theinhibitor concentration using the GraphPad Prism 5.0 software

To measure the effect of test compounds on the level of pBAD in cells inthe presence of human whole blood, human heparinized whole blood(Biological Specialty Corp, Colmar Pa.) was obtained and 350 μL/well wasadded to a 96-well 2 mL block plate (Costar 3961) in the presence orabsence of 20 μL of a concentration range of test compound(s). KMS12BMcells (1×10⁶) or MOLM-16 cell (5×10⁵) (DSMZ, Germany) in 25 μL of RPMIand 10% FBS (GIBCO) were added to each well. After 2.5 hours at 37° C.and 5% CO₂, red blood cells were lysed with erythrocyte lysis buffer(Qiagen) and the remaining cells were centrifuged at 1200 RPM. Theresulting pellets were lysed with 100 μL of cell extraction buffer (CellSignaling Technology) containing Halt, PMSF, and protease inhibitors(Thermo, Calbiochem, Sigma). The level of pBAD protein in the celllysates were then quantified in a commercial Human pBAD S112 ELISA Kit(Cell Signaling Technology). IC₅₀ determination was performed by fittingthe curve of percent inhibition versus the log of the inhibitorconcentration using the GraphPad Prism 5.0 software.

Example C Assay Data

Both Compound 1 phosphoric acid salt and Compound 1 dihydrochloric acidsalt were tested in the above-described assays of Examples A and B. Datais provided below in Table 30.

TABLE 30 Assay Type IC₅₀ (nM) IC₅₀ (nM) (Example No., Compound 1Compound 1 cell type) di-HCl salt H3PO4 salt PIM1 Enzyme (Ex. A) <35 <35PIM2 Enzyme (Ex. A) <35 <35 PIM3 Enzyme (Ex. A) <35 <35 Tumor cellproliferation <100 <100 (Ex. B, KMS12BM cells) Tumor cell proliferation<35 <35 (Ex. B, MOLM16 cells) pBAD KMS12BM cells (Ex. B) <35 <35 pBADwhole blood (Ex. B, <200 <200 KMS12BM cells) pBAD whole blood (Ex. B,<100 <100 MOLM16 cells)

Various modifications of the invention, in addition to those describedherein, will be apparent to those skilled in the art from the foregoingdescription. Such modifications are also intended to fall within thescope of the appended claims. Each reference, including withoutlimitation all patent, patent applications, and publications, cited inthe present application is incorporated herein by reference in itsentirety.

1-25. (canceled)
 26. A salt which is a hydrochloric acid salt ofN-{(7R)-4-[(3R,4R,5S)-3-amino-4-hydroxy-5-methylpiperidin-1-yl]-7-hydroxy-6,7-dihydro-5H-cyclopenta[b]pyridin-3-yl}-6-(2,6-difluorophenyl)-5-fluoropyridine-2-carboxamide.27. The salt of claim 26, wherein the salt is a dihydrochloric acidsalt.
 28. A solid form of the salt of claim
 27. 29. The solid form ofclaim 28 which is crystalline.
 30. The solid form of claim 29 having anXRPD pattern substantially as shown in FIG.
 4. 31. The solid form ofclaim 29 having a melting point of about 213° C.
 32. The salt of claim26, wherein the salt is a monohydrochloric acid salt.
 33. A solid formof the salt of claim
 32. 34. The solid form of claim 33 which iscrystalline.
 35. The solid form of claim 34 having an XRPD patternsubstantially as shown in FIG.
 22. 36. The solid form of claim 34 havinga melting point of about 209° C. 37-101. (canceled)
 102. The solid formof claim 29 having a DSC thermogram substantially as shown in FIG. 5.103. The solid form of claim 29 having a TGA thermogram substantially asshown in FIG.
 6. 104. The solid form of claim 29 having at least onecharacteristic XRPD peak, in terms of 2-theta, selected from about 8.3,about 18.9, and about 25.0 degrees.
 105. The solid form of claim 29having two or more characteristic XRPD peak, in terms of 2-theta,selected from about 8.3, about 18.9, and about 25.0 degrees.
 106. Thesolid form of claim 34 having a DSC thermogram substantially as shown inFIG.
 23. 107. The solid form of claim 34 having at least onecharacteristic XRPD peak, in terms of 2-theta, selected from about 7.8,about 8.8, about 12.6, about 14.5, about 17.4, about 23.8, and about25.2 degrees.
 108. The solid form of claim 34 having two or morecharacteristic XRPD peak, in terms of 2-theta, selected from about 7.8,about 8.8, about 12.6, about 14.5, about 17.4, about 23.8, and about25.2 degrees.
 109. The solid form of claim 34 having three or morecharacteristic XRPD peak, in terms of 2-theta, selected from about 7.8,about 8.8, about 12.6, about 14.5, about 17.4, about 23.8, and about25.2 degrees.